1 00:00:05,920 --> 00:00:07,400 >>WELCOME TO THE WEDNESDAY 2 00:00:07,400 --> 00:00:11,400 AFTERNOON LECTURE SERIES. THIS 3 00:00:11,400 --> 00:00:15,840 IS OF COURSE THE NIH-WIDE FORUM 4 00:00:15,840 --> 00:00:17,600 FOR THE MOST DISTINGUISHED 5 00:00:17,600 --> 00:00:21,960 SCIENTISTST IN BIOMEDICAL 6 00:00:21,960 --> 00:00:29,520 RESEARCH. I'LL BE HOSTING THE 7 00:00:29,520 --> 00:00:31,680 SPEAKER TODAY. HAU WU FROM THE 8 00:00:31,680 --> 00:00:33,960 FIELD OF STRUCTURAL BIOLOGY AND 9 00:00:33,960 --> 00:00:34,440 VISITING FROM HARVARD MEDICAL SCHOOL. 10 00:00:34,440 --> 00:00:35,520 DR. HAO WU IS A PROFESSOR 11 00:00:35,520 --> 00:00:44,920 AT HARVARD AND ALSO AN ASSOCIATE 12 00:00:44,920 --> 00:00:46,800 DIRECTOR AT THE BOSTON 13 00:00:46,800 --> 00:00:49,960 CHILDREN'S HOSPITAL AND WORLD 14 00:00:49,960 --> 00:00:55,760 RENOWNED FOR HER WORK ON SIGNAL 15 00:00:55,760 --> 00:00:58,000 INSTRUCTION IN CELL DEATH. SHE 16 00:00:58,000 --> 00:01:01,840 EARNED HER BACHELOR'S DEATH IN 17 00:01:01,840 --> 00:01:05,680 BEIJING, CHINA AND THEN WENT TO 18 00:01:05,680 --> 00:01:07,200 PEKING UNION MEDICAL COLLEGE, 19 00:01:07,200 --> 00:01:09,200 WHILE AT COLLEGE SHE 20 00:01:09,200 --> 00:01:09,960 PARTICIPATED IN LABORATORY 21 00:01:09,960 --> 00:01:13,400 RESEARCH AND IMMUNOLOGY AND AS 22 00:01:13,400 --> 00:01:19,400 RUMOR WOULD HAVE IT 23 00:01:19,400 --> 00:01:24,560 WITH RESEARCH AND 24 00:01:24,560 --> 00:01:27,080 RECEIVED A PH.D. FROM PURDUE 25 00:01:27,080 --> 00:01:29,120 UNIVERSITY. HER POSTDOC WAS AT 26 00:01:29,120 --> 00:01:31,160 COLUMBIA UNIVERSITY WITH ANOTHER 27 00:01:31,160 --> 00:01:34,280 RENOWNED STRUCTURAL BIOLOGIST 28 00:01:34,280 --> 00:01:37,360 WAYNE HENDRICKSON AND IN 1997 29 00:01:37,360 --> 00:01:40,240 BECAME A FACULTY MEMBER AT THE 30 00:01:40,240 --> 00:01:42,480 COLLEGE OF MEDICINE IN NEW YORK 31 00:01:42,480 --> 00:01:43,800 CITY. SHE WAS THERE FOR FIFTEEN 32 00:01:43,800 --> 00:01:49,320 YEARS AND MOVED HER LAB 33 00:01:49,320 --> 00:01:51,160 NORTHWARD TO HARVARD. AMONG HER 34 00:01:51,160 --> 00:01:53,280 MANY, MANY LANDMARK DISCOVERS 35 00:01:53,280 --> 00:01:55,880 AND I CAN SAY THIS FROM PERSONAL 36 00:01:55,880 --> 00:01:58,720 EXPERIENCE BECAUSE WE HAD THE 37 00:01:58,720 --> 00:02:01,840 GOOD FORTUNE TO COLLABORATE WITH 38 00:02:01,840 --> 00:02:04,040 HER. WAS IDENTIFYING THE 39 00:02:04,040 --> 00:02:05,840 ASSEMBLIES OF VARIOUS PROTEINS 40 00:02:05,840 --> 00:02:07,280 WITHIN THE CELL THAT CARRY OUT 41 00:02:07,280 --> 00:02:08,880 IMPORTANT FUNCTIONS IN CELL 42 00:02:08,880 --> 00:02:10,200 DEATH AND IMMUNITY. 43 00:02:10,200 --> 00:02:11,800 PARTICULARLY THE PROTEINS THAT 44 00:02:11,800 --> 00:02:13,920 WE HAD A STRONG INTEREST IN FROM 45 00:02:13,920 --> 00:02:15,880 A PUBLIC HEALTH AND DISEASE 46 00:02:15,880 --> 00:02:17,280 STANDPOINT I WOULD SAY SHE 47 00:02:17,280 --> 00:02:19,480 REWROTE THE TEST BOOKS WHEN SHE 48 00:02:19,480 --> 00:02:22,400 DISCOVERED THE INFLAM MA SOEM 49 00:02:22,400 --> 00:02:24,880 ACTIVATION THROUGH THESE HIGHER 50 00:02:24,880 --> 00:02:25,840 ORDER ASSEMBLIES. 51 00:02:25,840 --> 00:02:27,120 A THEME THAT GOES ON THROUGHOUT 52 00:02:27,120 --> 00:02:28,800 HER WORK AND ON MANY DIFFERENT 53 00:02:28,800 --> 00:02:31,160 IMPORTANT ASPECTS OF THE IMMUNE 54 00:02:31,160 --> 00:02:33,520 SYSTEM. SHE FOUND THAT THESE 55 00:02:33,520 --> 00:02:35,800 DEBT DOMAIN COMPLEXES HAVE A 56 00:02:35,800 --> 00:02:37,760 HELIO SYMMETRY THAT HARKENS BACK 57 00:02:37,760 --> 00:02:41,120 TO WORK BY FRANCIS AND OTHER 58 00:02:41,120 --> 00:02:43,120 OUTSTANDING BIOLOGISTS AND HAVE 59 00:02:43,120 --> 00:02:45,040 UNUSUAL STOIC I DON'T MEAN 60 00:02:45,040 --> 00:02:50,560 TRIES. 5-7 FOR THE PIDD, RAIDD 61 00:02:50,560 --> 00:02:55,160 COMPLEX FOR BASE TWO ACTIVATION. 62 00:02:55,160 --> 00:02:58,240 AND IN THE CAS BAIT 8 63 00:02:58,240 --> 00:03:00,480 ACTIVATION. THE TNF REACCEPT 64 00:03:00,480 --> 00:03:06,280 FORFAMILY AND 644 MORE MID 8. 65 00:03:06,280 --> 00:03:08,600 THESE ARE VERY UNUSUAL 66 00:03:08,600 --> 00:03:10,280 STRUCTURES I DON'T KNOW IF THOSE 67 00:03:10,280 --> 00:03:14,680 ARE FIBONACCI NUMBERS OR WHY 68 00:03:14,680 --> 00:03:17,160 NATURE EVOLVED TO HAVE THOSE 69 00:03:17,160 --> 00:03:18,360 NUMBERS BUT CAME DIRECTLY FROM 70 00:03:18,360 --> 00:03:21,720 HER INSIGHTS. HER LAB HAS DONE 71 00:03:21,720 --> 00:03:24,800 OTHER THINGS. THE HALLMARK OF 72 00:03:24,800 --> 00:03:27,360 ALZHEIMER'S DISEASE AND HAS 73 00:03:27,360 --> 00:03:30,240 BECOME AN OUTSTANDING CRY OWE 74 00:03:30,240 --> 00:03:31,560 EM. SCIENTIST IN ADDITION TO 75 00:03:31,560 --> 00:03:33,920 HER WORK IN CRYSTALLOGRAPHY SO 76 00:03:33,920 --> 00:03:36,160 NOW SHE'S IMAGING EVEN LARGER 77 00:03:36,160 --> 00:03:37,440 STRUCTURES IN THE CELL AND I 78 00:03:37,440 --> 00:03:38,680 JUST WANT TO POINT OUT THIS IS 79 00:03:38,680 --> 00:03:40,800 JUST A SNAPSHOT OF HER 80 00:03:40,800 --> 00:03:43,160 REMARKABLE CAREER. SHE'S HAD 81 00:03:43,160 --> 00:03:45,280 MANY, MANY HONORS FOR THE 82 00:03:45,280 --> 00:03:46,640 CONTRIBUTIONS SHE'S MADE. SHE 83 00:03:46,640 --> 00:03:48,680 WAS RECIPIENT OF THE MAYOR'S 84 00:03:48,680 --> 00:03:51,040 AWARD FROM NEW YORK CITY AND I 85 00:03:51,040 --> 00:03:53,160 ASSUME YOU GOT THAT FROM MIKE 86 00:03:53,160 --> 00:03:56,960 BLOOM B 87 00:03:56,960 --> 00:03:57,960 BLOOMBERG. THIS YEAR SHE WON AN 88 00:03:57,960 --> 00:04:02,400 AWARD FROM THE ASSOCIATION FROM 89 00:04:02,400 --> 00:04:03,040 IMMUNOLOGISTS AND ALSO ELECTED 90 00:04:03,040 --> 00:04:04,120 FOR THE ADVANCEMENT OF SCIENCES 91 00:04:04,120 --> 00:04:05,840 AND THE NATIONAL ACADEMY OF 92 00:04:05,840 --> 00:04:08,240 SCIENCES AND FROM US RECEIVED 93 00:04:08,240 --> 00:04:10,440 THE PRESTIGIOUS PIONEER AWARD 94 00:04:10,440 --> 00:04:12,040 GRANT WHICH IS GIVEN TO VERY FEW 95 00:04:12,040 --> 00:04:14,720 OF THE OUTSTANDING SCIENTISTS IN 96 00:04:14,720 --> 00:04:16,360 THE UNITED STATES AND IT MUST 97 00:04:16,360 --> 00:04:19,120 MAKE HER FEEL VERY GOOD TO BE AN 98 00:04:19,120 --> 00:04:21,120 HONOR PROFESSOR IN PEKING UNION 99 00:04:21,120 --> 00:04:27,120 MEDICAL COLLEGE, HER ALMA MATER 100 00:04:27,120 --> 00:04:32,960 AND AS WELL AN AN HONORARY 101 00:04:32,960 --> 00:04:33,640 PROFESSOR. THANK YOU FOR BEING 102 00:04:33,640 --> 00:04:34,520 HERE WITH US TODAY. AND I 103 00:04:34,520 --> 00:04:35,960 WELCOME YOU TO NIH. 104 00:04:35,960 --> 00:04:38,440 >> (APPLAUSE). 105 00:04:38,440 --> 00:04:42,400 >> THANK YOU SO MUCH MIKE, THAT 106 00:04:42,400 --> 00:04:44,360 WAS VERY KIND OF YOU. 107 00:04:44,360 --> 00:04:47,440 I'M DELIGHTED 108 00:04:47,440 --> 00:04:49,240 TO BE HERE. ACTUALLY I HAD GREAT 109 00:04:49,240 --> 00:04:51,760 FUN TALKING TO SOME OF YOU 110 00:04:51,760 --> 00:04:54,680 TODAY. THIS MORNING WITH PIS AS 111 00:04:54,680 --> 00:04:57,880 WELL AS STUDENTS AND POSTDOCS. 112 00:04:57,880 --> 00:05:00,720 SO I WOULD LIKE TO SPEND THE 113 00:05:00,720 --> 00:05:02,360 TIME TODAY TO TELL YOU SOMETHING 114 00:05:02,360 --> 00:05:04,600 THAT WE HAVE BEEN WORKING ON FOR 115 00:05:04,600 --> 00:05:06,080 A LONG TIME BUT NOW JUST FINALLY 116 00:05:06,080 --> 00:05:08,480 THEY ARE COMING TOGETHER AS WELL 117 00:05:08,480 --> 00:05:10,880 AS SOME UNPUBLISHED DATA. AND 118 00:05:10,880 --> 00:05:12,520 GUESS WHO I BUMP INTO THIS 119 00:05:12,520 --> 00:05:16,520 MORNING? TRYING TO GET 120 00:05:16,520 --> 00:05:20,720 BREAKFAST. SO HERE ARE MY 121 00:05:20,720 --> 00:05:22,760 DISCLOSURES. SO LET ME GET 122 00:05:22,760 --> 00:05:26,560 RIGHT TO IT. SO INFLAMMASOMES 123 00:05:26,560 --> 00:05:28,520 WERE FIRST IDENTIFIED, JUST WANT 124 00:05:28,520 --> 00:05:30,440 TO MAKE SURE THIS MOUSE WORKS. 125 00:05:30,440 --> 00:05:35,440 YEAH, OH. IS THIS THE PEN? 126 00:05:35,440 --> 00:05:40,920 OKAY. IDENTIFIED AS THE SUPER 127 00:05:40,920 --> 00:05:45,120 MOLECULAR COMPLEX THAT ACTIVATE 128 00:05:45,120 --> 00:05:55,960 ONE AND THIS CAME FROM YOURJURG 129 00:06:01,240 --> 00:06:04,640 LAB. THIS IS A PC, IT'S NOT A 130 00:06:04,640 --> 00:06:08,160 MAC, OKAY. THE FIRST INFLAM 131 00:06:08,160 --> 00:06:08,920 SOME WAS ACTUALLY IDENTIFIED IN 132 00:06:08,920 --> 00:06:12,760 MY FRIEND AND COLLABORATOR'S 133 00:06:12,760 --> 00:06:14,680 LAB. THE LATE JURG AS YOU CAN 134 00:06:14,680 --> 00:06:17,320 SEE FROM THE BEGINNING IT IS 135 00:06:17,320 --> 00:06:19,920 ALREADY KNOWN THAT INFLAM SOMES 136 00:06:19,920 --> 00:06:21,120 CONTAIN A PROSEEN NOPE AS THE 137 00:06:21,120 --> 00:06:23,360 SENSOR AND THEN THE ADAPT TOR 138 00:06:23,360 --> 00:06:25,120 AND THEN OF COURSE CASPASE ONE 139 00:06:25,120 --> 00:06:27,520 BECAUSE THAT'S THE FUNCTION FOR 140 00:06:27,520 --> 00:06:28,760 THE INFLAMMASOMES AND THE FACT 141 00:06:28,760 --> 00:06:30,120 THAT THEY FORM THESE LARGE KNOW 142 00:06:30,120 --> 00:06:31,520 LEB YOU WHAT ARE ASSEMBLIES AS 143 00:06:31,520 --> 00:06:33,400 YOU CAN SEE FROM THEIR MIGRATION 144 00:06:33,400 --> 00:06:35,280 FROM THEIR COLUMN. 145 00:06:35,280 --> 00:06:39,840 SO AS YOU CAN SEE, FROM THIS 146 00:06:39,840 --> 00:06:41,440 TIMELINE, THE INFLAMMASOME 147 00:06:41,440 --> 00:06:44,160 DISCOVERY REALLY WAS A 148 00:06:44,160 --> 00:06:45,440 BREAKTHROUGH FOR THE FIELD OF 149 00:06:45,440 --> 00:06:47,040 FEVER AND INFLAMMATION BECAUSE 150 00:06:47,040 --> 00:06:51,080 IT FOLLOWS THE FIRST DISCOVERY 151 00:06:51,080 --> 00:06:56,400 OF THE INTERLIEU ONE-1 CYTOKINE 152 00:06:56,400 --> 00:07:01,720 AND THEN THE CASPASE-1 THAT 153 00:07:01,720 --> 00:07:05,400 CLEAVES. SO NOW OF COURSE, 154 00:07:05,400 --> 00:07:08,800 INFLAMMASOME ACTIVATE THIS 155 00:07:08,800 --> 00:07:11,440 CASPASE-1. WE NOW KNOW THAT 156 00:07:11,440 --> 00:07:12,920 INFLAMMASOMES ARE COMPOSED OF 157 00:07:12,920 --> 00:07:14,960 THESE SUPERMOLECULAR STRUCTURES 158 00:07:14,960 --> 00:07:16,680 AND I LIST HERE SOME OF THE 159 00:07:16,680 --> 00:07:20,040 SENSOR PROTEINS THAT'S IN THESE 160 00:07:20,040 --> 00:07:21,960 INFLAMMASOMES AND IN TODAY'S 161 00:07:21,960 --> 00:07:25,280 TALK I WILL TALK ABOUT SOME OF 162 00:07:25,280 --> 00:07:27,840 OUR WORK ON THE INFLAMMASOME, AT 163 00:07:27,840 --> 00:07:29,160 LEAST THAT WILL BE PART OF THE 164 00:07:29,160 --> 00:07:31,920 FOCUS OF THE TALK. SO THESE 165 00:07:31,920 --> 00:07:35,080 SENSOR PROTEINS UPON ACTIVATION 166 00:07:35,080 --> 00:07:39,560 BY PATHOGENIC OR ENDOGENOUS 167 00:07:39,560 --> 00:07:41,880 DANGER SIGNALS, IT COULD ADAPT A 168 00:07:41,880 --> 00:07:44,800 PROTEIN AND THEN CASPASE-1 OF 169 00:07:44,800 --> 00:07:46,840 COURSE AND THIS WILL GENERATE 170 00:07:46,840 --> 00:07:51,960 AUTO PROCESSING OF CASPASE-1 TO 171 00:07:51,960 --> 00:07:53,200 INDUCE INFLAMMASOME ACTIVATION 172 00:07:53,200 --> 00:07:56,520 SO THE INFLAMMASOME ITSELF OFTEN 173 00:07:56,520 --> 00:07:59,760 FORMS AT A SINGLE SPEC IN THE 174 00:07:59,760 --> 00:08:01,720 CELL. SO THIS I'LL COME BACK 175 00:08:01,720 --> 00:08:02,320 LATER. THIS IS A SPECIAL 176 00:08:02,320 --> 00:08:06,840 LOCATION IN THE CELL. AT THE 177 00:08:06,840 --> 00:08:08,840 MICROTUBULE ORGANIZING CENTER SO 178 00:08:08,840 --> 00:08:11,480 ACTIVATING CASPASE-1 OR OTHER 179 00:08:11,480 --> 00:08:13,400 CASPASES THAT ARE ALSO 180 00:08:13,400 --> 00:08:15,400 PROINFLAMMATORY WHICH ARE 181 00:08:15,400 --> 00:08:17,120 INDUCED TO ACTIVATE THROUGH WHAT 182 00:08:17,120 --> 00:08:21,440 WE CALL A NON CANONICAL 183 00:08:21,440 --> 00:08:23,400 INFLAMMASOME PATHWAY CAN ALL 184 00:08:23,400 --> 00:08:27,320 CLEAVE THIS PROTEIN KNOWN AT GAS 185 00:08:27,320 --> 00:08:30,320 DWER PIN D INITIALLY DISCOVERED 186 00:08:30,320 --> 00:08:33,000 IN THE LAB. FROM ONE DAY USED 187 00:08:33,000 --> 00:08:34,320 GENETIC APPROACHES TO TRY TO 188 00:08:34,320 --> 00:08:38,560 LOOK FOR PROTEINS INVOLVED IN 189 00:08:38,560 --> 00:08:41,400 PAR RIP TO SIS. SO IT TURNS OUT 190 00:08:41,400 --> 00:08:44,920 IT CAN BE CLEAVED BY THESE 191 00:08:44,920 --> 00:08:46,440 CASPASES AND THEN THE 192 00:08:46,440 --> 00:08:47,680 INDETERMINABLE REGION THAT'S 193 00:08:47,680 --> 00:08:49,440 RELEASED FROM THIS CLEAVAGE AS 194 00:08:49,440 --> 00:08:52,920 WE SHOWED AND SEVERAL OTHER LABS 195 00:08:52,920 --> 00:08:54,880 SHOWED INTERACT WITH LIPIDS AT 196 00:08:54,880 --> 00:08:57,840 THE CELL MEMBRANE AND THEN FORM 197 00:08:57,840 --> 00:08:58,880 PORES AND THESE PORES ARE 198 00:08:58,880 --> 00:09:01,440 IMPORTANT FOR THE RELEASE OF THE 199 00:09:01,440 --> 00:09:03,640 MATERIAL CYTOKINE TO THE OUTSIDE 200 00:09:03,640 --> 00:09:06,440 OF THE CELL BECAUSE THESE MATURE 201 00:09:06,440 --> 00:09:08,960 CYTOKINES DO NOT HAVE A SIGNAL 202 00:09:08,960 --> 00:09:10,560 SEQUENCE AS WELL TO INDUCE THE 203 00:09:10,560 --> 00:09:13,000 CELL DEATH THAT'S HIGHLY 204 00:09:13,000 --> 00:09:14,240 INFLAMMATORY KNOWN AS PIE ROP TO 205 00:09:14,240 --> 00:09:17,240 SIS. SO LET ME START FROM THE 206 00:09:17,240 --> 00:09:21,920 NRP 3 INFLAMMASOME. A LITTLE 207 00:09:21,920 --> 00:09:25,440 BIT DIFFERENT FROM OTHER IMMUNE 208 00:09:25,440 --> 00:09:27,640 SENSORS. NRP 3 IS NOT A 209 00:09:27,640 --> 00:09:29,440 RECOGNITION RECEPTOR. INSTEAD 210 00:09:29,440 --> 00:09:31,160 IT SENSES SOMETHING MORE GENERAL 211 00:09:31,160 --> 00:09:33,440 AND THE EMERGING CONCEPT IS THAT 212 00:09:33,440 --> 00:09:37,800 NRP 3 SENSES SOME KIND OF A CELL 213 00:09:37,800 --> 00:09:39,560 MEMBRANE DAMAGE. SO MANY 214 00:09:39,560 --> 00:09:42,240 DIFFERENT THINGS CAN ACTIVATE 215 00:09:42,240 --> 00:09:46,200 NLRP 3. I HIGHLIGHT A COUPLE 216 00:09:46,200 --> 00:09:46,920 HERE. 217 00:09:46,920 --> 00:09:52,560 ONE IS THE BACTERIAL TOXIN NIGER 218 00:09:52,560 --> 00:09:54,800 RICIN BECAUSE IT ENDANGERS THE 219 00:09:54,800 --> 00:09:58,240 CELL MEMBRANE OR EXTRACELLULAR 220 00:09:58,240 --> 00:10:01,240 ATP WHICH IS A SIGNAL THAT CAN 221 00:10:01,240 --> 00:10:02,640 ACTIVATE A CHANNEL. AND THIS 222 00:10:02,640 --> 00:10:05,440 PARTICULAR STRUCTURES IN A CELL 223 00:10:05,440 --> 00:10:08,800 THAT CAN EACH INDUCE OR 224 00:10:08,800 --> 00:10:09,680 ASSOCIATE IT WITH CERTAIN 225 00:10:09,680 --> 00:10:13,440 IMPORTANT HUMAN DISEASES SUCH AS 226 00:10:13,440 --> 00:10:19,680 YOUR RISK ACID FOR GOUT. 227 00:10:19,680 --> 00:10:22,800 CHOLESTEROL CRYSTALS AND EVEN MP 228 00:10:22,800 --> 00:10:26,400 2.5 FOR ENVIRONMENTAL LAWN 229 00:10:26,400 --> 00:10:30,040 DISEASES. SO NLRP 3 IF YOU LOOK 230 00:10:30,040 --> 00:10:33,680 AT ITS DOMAIN OPTIMIZATION HAS 231 00:10:33,680 --> 00:10:37,440 THIS POWER DOMAIN AND THIS 232 00:10:37,440 --> 00:10:39,040 INDETERMINATES IS PART OF THE 233 00:10:39,040 --> 00:10:42,600 DOMAIN FAMILY AND THEN IT HAS 234 00:10:42,600 --> 00:10:44,760 THE SO-CALLED DOMAIN THAT IS 235 00:10:44,760 --> 00:10:45,480 ESSENTIALLY ATPASE BUT IT'S 236 00:10:45,480 --> 00:10:47,760 FUNCTION IS NOT TO GENERATE A 237 00:10:47,760 --> 00:10:50,680 FORCE, FOR EXAMPLE, DUE TO APT 238 00:10:50,680 --> 00:10:52,760 HYDROLYSIS BUT RATHER, BY 239 00:10:52,760 --> 00:10:57,320 BINDING TO ATP SOMEHOW INDUCES 240 00:10:57,320 --> 00:10:59,520 AGGREGATION OF THE PROTEIN IN 241 00:10:59,520 --> 00:11:01,400 ORDER TO PERFORM ITS FUNCTION 242 00:11:01,400 --> 00:11:05,080 AND THE DOMAIN IS WELL-KNOWN FOR 243 00:11:05,080 --> 00:11:08,400 THE FUNCTION. THE AC ADAPTER 244 00:11:08,400 --> 00:11:12,000 ALSO HAS A PYD PROTEIN AND THEY 245 00:11:12,000 --> 00:11:14,000 CAN INTERACT THROUGH THESE 246 00:11:14,000 --> 00:11:16,240 STRUCTURES THAT IS COMMON TO THE 247 00:11:16,240 --> 00:11:19,920 ENTIRE DOMAIN FAMILY. CARD IS 248 00:11:19,920 --> 00:11:24,360 ALSO A FAMILY MEMBER. AND ALSO 249 00:11:24,360 --> 00:11:28,160 ABLE TO RECRUIT THE CASPASE 250 00:11:28,160 --> 00:11:29,400 MOLECULES WHICH THEN BRING THE 251 00:11:29,400 --> 00:11:31,680 CASPASE TOGETHER FOR THEIR 252 00:11:31,680 --> 00:11:33,720 ACTIVATION. SO INTERESTINGLY IN 253 00:11:33,720 --> 00:11:35,480 CELLS, THEY FORM THESE LARGE 254 00:11:35,480 --> 00:11:37,440 SPECS, THAT ARE ESSENTIALLY 255 00:11:37,440 --> 00:11:43,760 LARGE AGGREGATES THAT CAN 256 00:11:43,760 --> 00:11:48,480 FURTHER KA CAPITULATE THEM IN VO 257 00:11:48,480 --> 00:11:52,760 SO THE QUESTION IS HOW DO THEN 258 00:11:52,760 --> 00:11:55,920 NLRP 3 FORM THE INITIAL PLATFORM 259 00:11:55,920 --> 00:11:58,840 IN ORDER TO KNEW COLLIUATE THE 260 00:11:58,840 --> 00:12:04,120 FORMATION OF THE PYD AND THE CA 261 00:12:04,120 --> 00:12:04,520 KRS 262 00:12:04,520 --> 00:12:05,800 CARD AND I WILL GO THROUGH THE 263 00:12:05,800 --> 00:12:07,760 JOURNEY THAT WE WENT THROUGH 264 00:12:07,760 --> 00:12:09,600 THROUGH THREE DIFFERENT 265 00:12:09,600 --> 00:12:12,440 STRUCTURES AND THIS EXTENSIVE 266 00:12:12,440 --> 00:12:14,000 INTRACELLULAR TRACKING THAT WE 267 00:12:14,000 --> 00:12:16,720 DISCOVERED ALONG THE WAY SO THE 268 00:12:16,720 --> 00:12:20,080 FIRST STRUCTURE THAT WE SOLVED 269 00:12:20,080 --> 00:12:25,400 WITH NLRP 3 IS THE NLPR 3 THAT 270 00:12:25,400 --> 00:12:29,360 DOESN'T HAVE THE PYD. WE WORRY 271 00:12:29,360 --> 00:12:30,760 THAT HAD THE ABILITY WOULD MAKE 272 00:12:30,760 --> 00:12:35,120 THIS PROTEIN HIGHLY AGGREGATE. 273 00:12:35,120 --> 00:12:37,160 SO WE SOLVED THIS STRUCTURE IN 274 00:12:37,160 --> 00:12:40,600 COMPLEX WITH NEK 7. WHAT IS NEK 275 00:12:40,600 --> 00:12:44,000 7? NEK 7 IN FACT IS A CENTRO 276 00:12:44,000 --> 00:12:47,480 SEW MALL KINASE. AND IT THE 277 00:12:47,480 --> 00:12:49,440 CATALYTIC ACTIVITY IS NOT 278 00:12:49,440 --> 00:12:53,520 REQUIRED FOR ITS SUPPORTING OF 279 00:12:53,520 --> 00:12:57,440 NRLP 3 ACTIVATION BUT THE 280 00:12:57,440 --> 00:12:59,400 SCAFFOLDING ACTIVATION THAT IS 281 00:12:59,400 --> 00:12:59,680 REQUIRED. 282 00:12:59,680 --> 00:13:03,840 WHEN WE SAW THE STRUCTURE WE SAW 283 00:13:03,840 --> 00:13:11,680 THE DOMAIN, DIRECT LY KO THIS NK 284 00:13:11,680 --> 00:13:17,000 7. WE ALSO KNOW THE NLRP 3 IS 285 00:13:17,000 --> 00:13:20,360 IN THE ADP BOUND STATE WHICH IS 286 00:13:20,360 --> 00:13:23,480 INACTIVE SO THEN A COUPLE YEARS 287 00:13:23,480 --> 00:13:27,120 AGO, WE SOLVED THE STRUCTURE OF 288 00:13:27,120 --> 00:13:29,880 THE RP 3 WITH THE PYD EVEN 289 00:13:29,880 --> 00:13:33,120 THOUGH THE PYD IS THERE IT LOOKS 290 00:13:33,120 --> 00:13:34,680 LIKE IT'S COMPLETELY BURIED 291 00:13:34,680 --> 00:13:38,160 WITHIN THIS -- WHAT WE CALL A 292 00:13:38,160 --> 00:13:40,200 DOUBLE RING CAPABLE. A CAPABLE 293 00:13:40,200 --> 00:13:43,440 THAT HAS TWO LAYERS. 506 ON TOP 294 00:13:43,440 --> 00:13:47,200 AND 506 ON THE BOTTOM AND PYD IS 295 00:13:47,200 --> 00:13:48,400 HIDDEN INSIDE THIS CAGE AND 296 00:13:48,400 --> 00:13:53,040 THAT'S PROBABLY WHY THAT THE 297 00:13:53,040 --> 00:13:54,840 NLRP 3 IS PROBABLY STILL 298 00:13:54,840 --> 00:14:00,880 INACTIVE AND THE NACHT DOMAIN IS 299 00:14:00,880 --> 00:14:03,840 STILL ADP BOUND NOT ATP BOUND. 300 00:14:03,840 --> 00:14:07,320 WHAT IS SURPRISING IS WHEN WE 301 00:14:07,320 --> 00:14:10,000 LOOKED AT THESE CAGES WE FOUND 302 00:14:10,000 --> 00:14:13,120 THEY'RE MOSTLY MEMBRANE BOUND SO 303 00:14:13,120 --> 00:14:15,160 WHEN WE FRACTION NATE THESE 304 00:14:15,160 --> 00:14:17,000 CELLS MOST OF THEM ARE IN THE 305 00:14:17,000 --> 00:14:19,200 MEMBRANE FACTION AND IF WE SOL 306 00:14:19,200 --> 00:14:20,840 YOU LOCALIZE THEM WE GET THE 307 00:14:20,840 --> 00:14:22,600 SAME KIND OF CAGE-LIKE STRUCTURE 308 00:14:22,600 --> 00:14:28,520 THAT WE SAW FROM THE SOLUBLE 309 00:14:28,520 --> 00:14:30,280 FRACTION. IN FACT THIS CAN BE 310 00:14:30,280 --> 00:14:31,720 CAPITULATED. IN THIS CASE, 311 00:14:31,720 --> 00:14:33,440 HERE, FOR EXAMPLE, IN THE WILD 312 00:14:33,440 --> 00:14:36,960 TYPE IMMORTALIZED BONE MARROW 313 00:14:36,960 --> 00:14:39,040 DERI 314 00:14:39,040 --> 00:14:40,680 DERIVED. THE FRANK HAS A BAND 315 00:14:40,680 --> 00:14:43,320 THAT CORRESPOND TO THE PURR FIE. 316 00:14:43,320 --> 00:14:44,120 WHERE THE PURIFIED WOULD HAVE TO 317 00:14:44,120 --> 00:14:49,160 RUN ON A NATIVE PAGE. 318 00:14:49,160 --> 00:14:51,800 SUGGESTING THESE MEMBRANES ARE 319 00:14:51,800 --> 00:14:53,040 CONTAINED IN THE CAGE SO THIS 320 00:14:53,040 --> 00:14:53,800 CAGE HAS THIS INTERESTING 321 00:14:53,800 --> 00:14:55,280 STRUCTURE THAT THIS REGION HERE, 322 00:14:55,280 --> 00:14:57,440 THIS REGION HERE, WHICH IS THE 323 00:14:57,440 --> 00:15:00,080 LINKER BETWEEN THE PYD AND THE 324 00:15:00,080 --> 00:15:01,560 MATCH DOMAIN RATE RIGHT HERE. 325 00:15:01,560 --> 00:15:04,120 THIS REGION HERE HAS A LOT OF 326 00:15:04,120 --> 00:15:05,480 POSITIVELY CHARGED RESIDUES. 327 00:15:05,480 --> 00:15:08,440 AND WE SHOW THAT WHEN YOU MUTATE 328 00:15:08,440 --> 00:15:09,280 THESE RESIDUES, THEY ACTUALLY 329 00:15:09,280 --> 00:15:12,760 CAN NO LONGER BE ON THE MEMBRANE 330 00:15:12,760 --> 00:15:17,400 AND THEY ALSO CANNOT FORM THE 331 00:15:17,400 --> 00:15:20,480 CAGE SUGGESTING THAT THIS 332 00:15:20,480 --> 00:15:22,160 BINDING TO THE MEMBRANE IS 333 00:15:22,160 --> 00:15:25,200 IMPORTANT FOR CAGE FORMATION. 334 00:15:25,200 --> 00:15:28,520 SO JAMES CHAN'S LAB A FEW YEARS 335 00:15:28,520 --> 00:15:31,640 AGO FOUND THAT THE STIMULATION 336 00:15:31,640 --> 00:15:35,480 INDUCES THIS. SO THE GOLGI 337 00:15:35,480 --> 00:15:37,440 WHICH USUALLY IS A CAP LIKE 338 00:15:37,440 --> 00:15:41,240 STRUCTURE RIGHT NEXT TO THE 339 00:15:41,240 --> 00:15:47,520 NUCLEUS IMPOUND THE INFLAMMASOME 340 00:15:47,520 --> 00:15:51,440 THE CAP COMPLETELY DISAAPPEAR 341 00:15:51,440 --> 00:15:53,400 AND BECOME VESICLES AND THEY GET 342 00:15:53,400 --> 00:15:55,760 TRAFFICKED TO THE MTOC. THIS IS 343 00:15:55,760 --> 00:16:02,440 AT THE MTOC AND THIS IS A TGN 344 00:16:02,440 --> 00:16:06,880 AND THESE ARE TRAFFICKED TO THE 345 00:16:06,880 --> 00:16:11,480 MTOC TO BRING NLRP 3 TO THE MTO 346 00:16:11,480 --> 00:16:13,280 KRVRJTS THIS IS A KINASE AND WE 347 00:16:13,280 --> 00:16:18,320 WILL COME BACK TO THAT IN A BIT 348 00:16:18,320 --> 00:16:19,880 THEN WE SHOW THE CAGE. WHY DO 349 00:16:19,880 --> 00:16:20,960 WE HAVE TO STRUCTURE. IT'S KIND 350 00:16:20,960 --> 00:16:23,520 OF A COMPLICATED STRUCTURE THAT 351 00:16:23,520 --> 00:16:25,400 PREVENTS ACTIVATION BUT WHY DO 352 00:16:25,400 --> 00:16:28,640 WE NEED IT FOR NLRP 3 ACTIVATION 353 00:16:28,640 --> 00:16:32,720 SO IT TURNED OUT THE CAGE ITSELF 354 00:16:32,720 --> 00:16:34,920 IS IMPORTANT FOR THE DISPERSION 355 00:16:34,920 --> 00:16:42,240 OF THE TGN AFTER NLRP 3 356 00:16:42,240 --> 00:16:44,800 STIMULATION. IN THE WILD TYPE 357 00:16:44,800 --> 00:16:51,040 MACROPHAGES, THESE ARE THE TGN 358 00:16:51,040 --> 00:16:53,040 STAINING. THESE ARE UPON 359 00:16:53,040 --> 00:16:57,320 STIMYOU LAYING WITH THE 360 00:16:57,320 --> 00:17:01,400 BACTERIAL TOXIN AND THE TGN THEY 361 00:17:01,400 --> 00:17:05,320 ALL ARE HERE AND DISPERSE INTO 362 00:17:05,320 --> 00:17:08,680 TINY VESICLES AND THIS 363 00:17:08,680 --> 00:17:12,160 DISPERSION IS DEPENDENT. IF YOU 364 00:17:12,160 --> 00:17:14,240 USE THE KNOCKOFF CELLS YOU DON'T 365 00:17:14,240 --> 00:17:15,880 GET THIS DISPERSION UPON 366 00:17:15,880 --> 00:17:18,480 TREATMENT WITHIN THE ACTIVATOR 367 00:17:18,480 --> 00:17:20,640 AND IF YOU RECONSTITUTE THE 368 00:17:20,640 --> 00:17:24,400 CELLS WITH A MUTANT OF NLRP 3 369 00:17:24,400 --> 00:17:27,120 THAT CANNOT FORM THE CAGE YOU 370 00:17:27,120 --> 00:17:30,880 ALSO DON'T GET TGN DISPERSION 371 00:17:30,880 --> 00:17:32,120 SUGGESTING THAT THE CAGE IS 372 00:17:32,120 --> 00:17:33,360 IMPORTANT FOR THIS PROCESS SO 373 00:17:33,360 --> 00:17:36,960 THIS LED US TO HYPOTHESIS THAT 374 00:17:36,960 --> 00:17:39,400 THE CAGE ITSELF MAKES THIS FOR A 375 00:17:39,400 --> 00:17:42,080 REASON. IT NOT ONLY SUPPRESSES 376 00:17:42,080 --> 00:17:45,440 THE NLRP 3 ACTIVATION BUT ALSO 377 00:17:45,440 --> 00:17:47,520 IMPORTANT FOR THIS EARLY STEP IN 378 00:17:47,520 --> 00:17:50,040 THIS ACTIVATION BY DISPERSING 379 00:17:50,040 --> 00:17:54,000 THE TGN SO NOW WE KEPT ON 380 00:17:54,000 --> 00:17:57,120 SOLVING NLRP 3 STRUCTURE IN THE 381 00:17:57,120 --> 00:17:58,880 CONFIRMATION. NO MATTER WHAT WE 382 00:17:58,880 --> 00:18:03,720 DO, EVERYTHING THAT WE PURIFY IS 383 00:18:03,720 --> 00:18:05,120 IN THE INACTIVE CONFIRMATION SO 384 00:18:05,120 --> 00:18:10,320 THIS CHANGED WHEN MY POSTDOC LE 385 00:18:10,320 --> 00:18:12,720 DECIDED TO TRY SOMETHING ELSE. 386 00:18:12,720 --> 00:18:21,760 HE COEXPRESSED NLRP 3 WITH NEK 387 00:18:21,760 --> 00:18:25,440 7. AND THEN HE TREATED THE 388 00:18:25,440 --> 00:18:28,800 CELLS WITH NIGH OWE RICIN, 389 00:18:28,800 --> 00:18:30,280 BASICALLY TREAT THE CELL WITH 390 00:18:30,280 --> 00:18:37,320 THE SAME STIMULUS AND THEN HE 391 00:18:37,320 --> 00:18:45,400 ADD THE ATP ANALOG. TO THIS TO 392 00:18:45,400 --> 00:18:46,520 STABILIZE THE CONFIRMATION AND 393 00:18:46,520 --> 00:18:51,240 ON TOP OF IT HE ADDED ASC 394 00:18:51,240 --> 00:18:52,480 BECAUSE IF WE HAVE -- IF WE 395 00:18:52,480 --> 00:18:54,400 DON'T HAVE ALL THREE OF THEM WE 396 00:18:54,400 --> 00:18:57,440 DO NOT GET THIS ORDERED 397 00:18:57,440 --> 00:18:59,920 STRUCTURE. I THINK PARTLY 398 00:18:59,920 --> 00:19:04,440 BECAUSE THE NLRP 3 INFLAMMASOME 399 00:19:04,440 --> 00:19:08,320 UNLIKE THE STRUCTURES WHICH WE 400 00:19:08,320 --> 00:19:11,560 SOLVED, IN THERE, ASC WAS NOT 401 00:19:11,560 --> 00:19:13,080 REQUIRED FOR THE COMPLEX BUT 402 00:19:13,080 --> 00:19:17,160 HERE YOU NEED THE ASC TO TABLIZE 403 00:19:17,160 --> 00:19:21,240 THE NLRP 3 COMPLEX. YOU CAN 404 00:19:21,240 --> 00:19:26,680 ONLY MAINTAIN IT BETWEEN NLRP 3, 405 00:19:26,680 --> 00:19:31,480 NEK 7 ANDS ASC AND YOU CAN SEE 406 00:19:31,480 --> 00:19:33,880 THE STRUCTURES THAT ARE EVEN 11 407 00:19:33,880 --> 00:19:35,560 FOLD OR 10 FOLD AND THE 408 00:19:35,560 --> 00:19:37,400 STRUCTURE WITH THE 10 FOLD REACH 409 00:19:37,400 --> 00:19:38,520 THE BEST RESOLUTION AND 410 00:19:38,520 --> 00:19:41,080 THEREFORE I'M SHOWING YOU THE 10 411 00:19:41,080 --> 00:19:44,880 FOLD STRUCTURE. SO THIS IS THE 412 00:19:44,880 --> 00:19:47,360 INDIVIDUAL SUBUNIT. IN THE MAP, 413 00:19:47,360 --> 00:19:49,840 THIS IS WHERE N EK 7 IS. SORRY, 414 00:19:49,840 --> 00:19:51,360 I'M NOT DRAWING IT WELL. THE 415 00:19:51,360 --> 00:19:53,520 YELLOW IS NEK 7. EVERYTHING 416 00:19:53,520 --> 00:19:56,600 ELSE IS NLRP 3. THIS WHOLE 417 00:19:56,600 --> 00:19:58,360 THING HERE AND THIS DOMAIN NOW 418 00:19:58,360 --> 00:20:00,600 HAS CHANGED THE CONFIRMATION TO 419 00:20:00,600 --> 00:20:03,240 AN ATP BOUND CONFIRMATION AND IF 420 00:20:03,240 --> 00:20:05,040 YOU LOOK AT THE SITE VIEW YOU 421 00:20:05,040 --> 00:20:10,040 CAN SEE THAT THE ADDED CYPD IS 422 00:20:10,040 --> 00:20:14,760 TOGETHER WITH THE -- NLPR 3 TO 423 00:20:14,760 --> 00:20:17,280 FORM A SHORT FILAMENT NEXT TO 424 00:20:17,280 --> 00:20:21,960 THE DISC. SO JUST SHOWING YOU 425 00:20:21,960 --> 00:20:28,320 BRIEFLY THE CONFIRMATION CHANGE 426 00:20:28,320 --> 00:20:31,280 THAT'S OCCURRING. BASICALLY 427 00:20:31,280 --> 00:20:33,040 THIS DOMAIN AS WELL AS PART OF 428 00:20:33,040 --> 00:20:34,720 THE ENACT DOMAIN TOGETHER HAVE 429 00:20:34,720 --> 00:20:36,480 THIS MASSIVE MOVEMENT. ALMOST 430 00:20:36,480 --> 00:20:38,640 90 DEGREE MOVEMENT AND I WON'T 431 00:20:38,640 --> 00:20:40,800 DETAIL THESE OTHER CHANGES BUT 432 00:20:40,800 --> 00:20:44,520 THERE'S ALSO LOCAL CHANGES IN 433 00:20:44,520 --> 00:20:47,080 OTHER IMPORTANT DOMAINS IN THE 434 00:20:47,080 --> 00:20:48,320 PROTEIN ESPECIALLY THIS DOMAIN 435 00:20:48,320 --> 00:20:58,440 THAT HAS BEEN IMPLICATED AS A 436 00:20:58,440 --> 00:21:00,440 SENSOR FOR INFLUX. I THINK 437 00:21:00,440 --> 00:21:02,320 THERE'S A LOT MORE STORY TO THAT 438 00:21:02,320 --> 00:21:04,080 BUT WE DON'T KNOW HOW THIS 439 00:21:04,080 --> 00:21:06,240 DOMAIN OR OTHER ASPECTS OF THE 440 00:21:06,240 --> 00:21:10,160 BIOLOGY SENSES THE ACTIVATORS SO 441 00:21:10,160 --> 00:21:13,280 HERE I JUST SHOW YOU THAT 442 00:21:13,280 --> 00:21:16,560 INSTEAD OF ATT -- ADP -- NOW WE 443 00:21:16,560 --> 00:21:21,000 HAVE ATP BOUND AND THIS RESIDUE 444 00:21:21,000 --> 00:21:23,600 HERE HISTIDINE 522 IN THE 445 00:21:23,600 --> 00:21:26,760 INACTIVE CONFIRMATION WOULD HAVE 446 00:21:26,760 --> 00:21:29,720 BEEN IN CLASH WITH THE ACTUAL 447 00:21:29,720 --> 00:21:32,360 GROUP IN THE ATP AND THEREFORE, 448 00:21:32,360 --> 00:21:35,760 THIS WOULD INDUCE A 449 00:21:35,760 --> 00:21:37,920 CONFIRMATIONAL CHANGE TO THE 450 00:21:37,920 --> 00:21:41,520 ACTIVE STATE WHEN ATP IS BOUND. 451 00:21:41,520 --> 00:21:42,800 SO MOST PROMINENT QUESTION FROM 452 00:21:42,800 --> 00:21:46,320 THE STRUCTURES I THINK IS THE 453 00:21:46,320 --> 00:21:48,440 FACT THAT NEK 7 WHICH I SHOWED 454 00:21:48,440 --> 00:21:50,160 YOU THIS BEFORE, ALREADY, NEK 7 455 00:21:50,160 --> 00:21:57,000 WHICH IS RIGHT HERE. IT DOESN'T 456 00:21:57,000 --> 00:21:59,520 REALLY ASSOCIATE WITH ANYTHING. 457 00:21:59,520 --> 00:22:02,360 I DOESN'T ALLEVIATE THIS AND THE 458 00:22:02,360 --> 00:22:03,960 QUESTION WOULD BE THEN WHAT IS 459 00:22:03,960 --> 00:22:09,400 THE FUNCTION OF NEK 7? SO WE 460 00:22:09,400 --> 00:22:12,600 HYPOTHESIZED BECAUSE NEK 7 ALSO 461 00:22:12,600 --> 00:22:13,920 BINDS THE LEUCINE RETREAT DOMAIN 462 00:22:13,920 --> 00:22:17,360 AND THIS IS THE SAME AS FOR THE 463 00:22:17,360 --> 00:22:19,760 LEUCINE RICH DOMAIN INTERACTION 464 00:22:19,760 --> 00:22:22,480 IN THE CAGE. SO WE HYPOTHESIZE 465 00:22:22,480 --> 00:22:24,080 THAT THESE TWO BINDINGS ARE 466 00:22:24,080 --> 00:22:25,160 COMPETING WITH EACH OTHER. AND 467 00:22:25,160 --> 00:22:27,280 INDEED IF WE START WITH A CAGE 468 00:22:27,280 --> 00:22:31,160 STRUCTURE AND THEN WE ADD NEK 7 469 00:22:31,160 --> 00:22:34,560 AS SHOWN HERE, THE CAGE WHICH IS 470 00:22:34,560 --> 00:22:35,520 THIS PEAK HERE AND THIS PEAK 471 00:22:35,520 --> 00:22:37,640 HERE THE CAGE WOULD BE DISRUPTED 472 00:22:37,640 --> 00:22:41,400 AND SUGGESTING THAT INDEED, NEK 473 00:22:41,400 --> 00:22:45,920 7 CAN DISRUPT A CAGE SO THIS 474 00:22:45,920 --> 00:22:49,200 BIOCHEMICAL DATA AS WELL AS CELL 475 00:22:49,200 --> 00:22:50,880 BIOLOGY AND STRUCTURAL DATA 476 00:22:50,880 --> 00:22:53,680 PROMPT US TO PROMOTE -- TO -- 477 00:22:53,680 --> 00:22:57,400 PROPOSE THIS HYPOTHESIS OF 478 00:22:57,400 --> 00:22:58,440 TRAFFICKING AND CONFIRMATIONAL 479 00:22:58,440 --> 00:23:04,080 CHANGES IN ACTIVATION. THAT IS, 480 00:23:04,080 --> 00:23:07,080 STARTING THERE MONOER HERRIC 481 00:23:07,080 --> 00:23:11,520 NLRP 3 OR THE CAGE FORM AT THE 482 00:23:11,520 --> 00:23:13,400 WORK. THERE MAY BE AN 483 00:23:13,400 --> 00:23:16,560 EQUILIBRIUM BETWEEN THIS STATE 484 00:23:16,560 --> 00:23:18,240 AND THE CAGE STATE. APPARENTLY 485 00:23:18,240 --> 00:23:21,400 IN NLRP 3 ACTIVATION WE IMAGINE 486 00:23:21,400 --> 00:23:25,400 THAT THE ATP EXCHANGE WOULD HAVE 487 00:23:25,400 --> 00:23:30,400 OCCURRED ALREADY SO THAT THIS 488 00:23:30,400 --> 00:23:32,320 NEK DOMAIN CHANGES CON FIR MAKE. 489 00:23:32,320 --> 00:23:34,600 THIS CLOSED CONFIRMATION IS MORE 490 00:23:34,600 --> 00:23:37,360 COMPACT WHEREAS THE ACTIVE ATP 491 00:23:37,360 --> 00:23:38,200 BOUND CONFIRMATION IS MORE 492 00:23:38,200 --> 00:23:41,520 STANDING UP LIKE THIS AND THEN 493 00:23:41,520 --> 00:23:44,480 THIS KIND OF CAGE IS ABLE TO BE 494 00:23:44,480 --> 00:23:48,440 TRANSPORTED TO THE MTOC ON THE 495 00:23:48,440 --> 00:23:51,560 MICROTUBULE NETWORK. THIS 496 00:23:51,560 --> 00:23:52,840 ACTUALLY BEEN KNOWN FOR UNITED 497 00:23:52,840 --> 00:23:56,040 STATES SOME TIME AND ONE OF THE 498 00:23:56,040 --> 00:23:59,520 FIST DISCOVERIES OF 499 00:23:59,520 --> 00:24:00,960 INFLAMMASOMES IS THAT IT IS 500 00:24:00,960 --> 00:24:05,120 SENSITIVE TO MICROTUBULE 501 00:24:05,120 --> 00:24:09,040 STRUCTURES AND IN FACT THIS IS 502 00:24:09,040 --> 00:24:11,520 AN INFLAMMASOME DRUG. ONCE IT 503 00:24:11,520 --> 00:24:14,840 GETS TRAFFICKED WE HYPOTHESIZE 504 00:24:14,840 --> 00:24:16,680 NEK 7 WOULD BE THERE TO DISRUPT 505 00:24:16,680 --> 00:24:21,400 THE CAGE AND BECAUSE THE TWO LAY 506 00:24:21,400 --> 00:24:23,160 NEVERS -- LAYERS ARE ALREADY 507 00:24:23,160 --> 00:24:25,040 LINKED TOGETHER. ONE CAN 508 00:24:25,040 --> 00:24:26,600 IMAGINE THAT THIS WILL JUST 509 00:24:26,600 --> 00:24:29,040 UNFURL. UNFURL THE CAGE INTO 510 00:24:29,040 --> 00:24:31,840 TWO FIVE SUBUNIT SUB COMPLEX. 511 00:24:31,840 --> 00:24:35,360 AND THEN THESE TWO CAN JUST COME 512 00:24:35,360 --> 00:24:37,200 TOGETHER TO FORM THE FINAL DISC 513 00:24:37,200 --> 00:24:39,320 THAT CONTAINS TEN SUBUNITS AND 514 00:24:39,320 --> 00:24:41,400 THIS OF COURSE NEED TO BE 515 00:24:41,400 --> 00:24:45,360 FURTHER STABILIZED BY THE BOUND 516 00:24:45,360 --> 00:24:51,440 ASC SHOWN HERE IN RED. WHY IT 517 00:24:51,440 --> 00:24:55,880 USE Z SUCH A COMPLEX ACTIVISM, 518 00:24:55,880 --> 00:25:04,480 WE THINK ANY INFLAMMASOME 519 00:25:04,480 --> 00:25:05,320 ACTIVITY. IF YOU PRODUCE DEATH 520 00:25:05,320 --> 00:25:08,000 THAT'S THE END OF THE CELL SO 521 00:25:08,000 --> 00:25:09,920 PROBABLY THE CELL HAVE DEVELOPED 522 00:25:09,920 --> 00:25:11,960 MANY DIFFERENT REGULATORY 523 00:25:11,960 --> 00:25:13,400 MECHANISM IN ORDER TO SUPPRESS 524 00:25:13,400 --> 00:25:15,920 THIS ACTIVATION UNTIL EVERY STEP 525 00:25:15,920 --> 00:25:18,640 IS CLICKED. EVERY STEP, EVERY 526 00:25:18,640 --> 00:25:23,560 CHECKPOINT IS CLICKED. SO THAT 527 00:25:23,560 --> 00:25:27,600 NLRP 3 CAN BE ACTIVATED. SO 528 00:25:27,600 --> 00:25:30,800 INTERESTINGLY NLRP 3 IS 529 00:25:30,800 --> 00:25:32,760 CONSERVED THROUGHOUT EVOLUTION 530 00:25:32,760 --> 00:25:34,440 AND BAT IS AN INTERESTING CASE. 531 00:25:34,440 --> 00:25:37,040 SO THE BAT IS KNOWN AS A 532 00:25:37,040 --> 00:25:38,880 RESERVOIR FOR MANY DIFFERENT 533 00:25:38,880 --> 00:25:41,400 VIRUSES AS YOU KNOW. WHICH MAY 534 00:25:41,400 --> 00:25:45,400 INCLUDE THE CORONAVIRUS. THE 535 00:25:45,400 --> 00:25:48,240 PANDEMIC THAT WE HAVE JUST 536 00:25:48,240 --> 00:25:54,960 SUFFERED THROUGH. SO IT TURNED 537 00:25:54,960 --> 00:25:57,280 OUT THEIR SAY SEW FORMS ARE 538 00:25:57,280 --> 00:25:58,840 DIFFERENT AND THIS MEANS IT IS 539 00:25:58,840 --> 00:25:59,120 DELETED. 540 00:25:59,120 --> 00:26:01,840 IF YOU LOOK AT THE DELETED 541 00:26:01,840 --> 00:26:04,640 ISOFORMS IT HAS -- WHICH IS 542 00:26:04,640 --> 00:26:07,720 ISOFORM 2 AND 4 IT HAS VERY LOW 543 00:26:07,720 --> 00:26:09,720 ACTIVITY AS IN 2 HERE AND 4 544 00:26:09,720 --> 00:26:15,200 HERE. VERY LOW ACTIVITY SO WE 545 00:26:15,200 --> 00:26:16,120 HYPOTHESIZED THAT BECAUSE OF 546 00:26:16,120 --> 00:26:18,840 THIS DELETION IN THE DOMAIN, IT 547 00:26:18,840 --> 00:26:20,360 STILL MAY BE ABLE TO FORM THE 548 00:26:20,360 --> 00:26:22,720 CAGE BUT MAY NOT BE ABLE TO BIND 549 00:26:22,720 --> 00:26:25,400 TO NEK 7 AND THEREFORE YOU 550 00:26:25,400 --> 00:26:34,440 CANNOT ACTIVATE THE BAD NLRP 3 551 00:26:34,440 --> 00:26:35,400 INFLAM SOME AND THIS IS 552 00:26:35,400 --> 00:26:36,520 SOMETHING WE'RE STUDYING. 553 00:26:36,520 --> 00:26:37,200 ANOTHER INTERESTING QUESTION 554 00:26:37,200 --> 00:26:41,040 THAT WE'RE TRYING TO ADDRESS IS 555 00:26:41,040 --> 00:26:45,080 WHAT ARE THE EFFECTS OF THESE 556 00:26:45,080 --> 00:26:49,240 CRY YOWE PYRIN ASSOCIATED 557 00:26:49,240 --> 00:26:50,600 PERIODIC SYNDROME MUTATIONS. 558 00:26:50,600 --> 00:26:52,880 ALMOST ALL THESE MUTATIONS ARE 559 00:26:52,880 --> 00:26:57,840 MAPPED TO THE NETWORKS -- NEK 560 00:26:57,840 --> 00:27:00,840 DOMAIN BUT WE DON'T KNOW WHETHER 561 00:27:00,840 --> 00:27:02,240 THIS AUTOMATIC CON FIR MAKAL 562 00:27:02,240 --> 00:27:04,800 CHANGE IN THESE PROTEINS WOULD 563 00:27:04,800 --> 00:27:06,280 ALREADY CAUSE DISPERSION OR YOU 564 00:27:06,280 --> 00:27:08,640 STILL NEED A SECOND STIMULUS, 565 00:27:08,640 --> 00:27:10,880 MAYBE A VERY WEAK STIMULUS IN 566 00:27:10,880 --> 00:27:11,840 ORDER TO ACTIVATE THE PATHWAY 567 00:27:11,840 --> 00:27:15,320 AND WE THOUGHT THIS COULD BE A 568 00:27:15,320 --> 00:27:18,640 VERY INTERESTING ONE TO ADDRESS. 569 00:27:18,640 --> 00:27:23,080 SO AS A SUMMARY FOR THIS PART OF 570 00:27:23,080 --> 00:27:28,520 THE TALK, BASICALLY, WE SHOW 571 00:27:28,520 --> 00:27:32,400 THAT HAD THE DIFFERENT NLRS. I 572 00:27:32,400 --> 00:27:36,360 TALKED ABOUT THE NLRP INFLAM 573 00:27:36,360 --> 00:27:37,400 SOME BUT THERE ARE OTHERS. WHAT 574 00:27:37,400 --> 00:27:41,160 WAS STRIKING TO ME WAS THAT THE 575 00:27:41,160 --> 00:27:43,160 DIFFERENT NLR PROTEINS ALL HAVE 576 00:27:43,160 --> 00:27:44,600 THE SAME PROTEIN ORGANIZATION. 577 00:27:44,600 --> 00:27:45,920 THEY ALL USE VERY DIFFERENT WAYS 578 00:27:45,920 --> 00:27:49,440 TO ACTIVATE THE INFLAMMASOME. 579 00:27:49,440 --> 00:27:54,280 FOR EXAMPLE I DIDN'T TALK ABOUT 580 00:27:54,280 --> 00:27:58,040 NLRP 6 BUT IT GOES INTO A PHASE 581 00:27:58,040 --> 00:28:00,680 SEPARATION. INTO DOUBLE 582 00:28:00,680 --> 00:28:02,400 STRANDED. AND THIS IS THE 583 00:28:02,400 --> 00:28:03,920 INITIAL SIGNALLING PLATFORM FOR 584 00:28:03,920 --> 00:28:08,280 THE NLRP 6 INFLAM IF A SOME 585 00:28:08,280 --> 00:28:09,440 ACTIVATION WHEREAS ONE USES A 586 00:28:09,440 --> 00:28:11,560 CLEAVAGE IN ITS PROTEIN IN ORDER 587 00:28:11,560 --> 00:28:16,840 TO RELEASE A TOXIC INFLAMMASOME 588 00:28:16,840 --> 00:28:19,840 FORMING FRAGMENT TO INDUCE THE 589 00:28:19,840 --> 00:28:22,040 ACTIVATION OF THIS PARTICULAR 590 00:28:22,040 --> 00:28:26,000 NLR INFLAMMASOME WE LOOK FORWARD 591 00:28:26,000 --> 00:28:28,440 TO LEARNING WHAT OTHER KIND OF 592 00:28:28,440 --> 00:28:31,120 INFLAMMASOMES MAY DO. IN ITS 593 00:28:31,120 --> 00:28:34,800 ABILITY TO INDUCE INFLAMMASOME 594 00:28:34,800 --> 00:28:38,600 FORMATION. SO NOW I WILL TALK 595 00:28:38,600 --> 00:28:45,200 ABOUT GSDMD IS A SUBSTRATE. WE 596 00:28:45,200 --> 00:28:48,800 ACTUALLY KNOW NOW THAT GSDMD CAN 597 00:28:48,800 --> 00:28:51,200 BE CLEAVED BUT OTHER ENZYMES I 598 00:28:51,200 --> 00:28:54,360 THINK I PROBABLY HAVE A SLIDE IN 599 00:28:54,360 --> 00:29:03,640 THE NEXT FEW SLIDES SO WE SHOWED 600 00:29:03,640 --> 00:29:06,000 THAT GASDERMIN BINDS. WHERE ARE 601 00:29:06,000 --> 00:29:11,120 THESE AT THE CELL? IN THE INNER 602 00:29:11,120 --> 00:29:14,200 LEAFLET AND THAT'S WHY GASDERMIN 603 00:29:14,200 --> 00:29:15,800 CAN PUNCH A HOLE FROM WITHIN AND 604 00:29:15,800 --> 00:29:17,400 ONCE THEY GET RELEASED FROM THE 605 00:29:17,400 --> 00:29:18,920 CELL THEY CANNOT ATTACK THE CELL 606 00:29:18,920 --> 00:29:20,400 ANYMORE BECAUSE THERE'S NO 607 00:29:20,400 --> 00:29:22,360 ACIDIC LIPIDS AND THE FACT THAT 608 00:29:22,360 --> 00:29:25,840 IT CAN ALSO INTERACT WITH 609 00:29:25,840 --> 00:29:28,720 CARDIOLIE PIN, THIS IS A LIPID 610 00:29:28,720 --> 00:29:31,680 THAT'S PRESENT IN MITOCHONDRIA 611 00:29:31,680 --> 00:29:33,400 AND ALSO SUGGESTS THAT 612 00:29:33,400 --> 00:29:35,320 GASDERMINS CAN DAMAGE THE 613 00:29:35,320 --> 00:29:36,200 MITOCHONDRIA AND THERE'S A LOT 614 00:29:36,200 --> 00:29:39,560 OF EVIDENCE SUPPORTING THAT. 615 00:29:39,560 --> 00:29:42,120 SO HERE'S A GASDERMIN FAMILY A 616 00:29:42,120 --> 00:29:44,560 FAMILY THAT HAS BEEN THERE FOR 617 00:29:44,560 --> 00:29:45,960 AWHILE. BUT DIDN'T DRAW A LOT 618 00:29:45,960 --> 00:29:49,400 OF ATTENTION TO THE FIELD. 619 00:29:49,400 --> 00:29:54,800 UNTIL THE DISCOVERY THAT THE 620 00:29:54,800 --> 00:29:56,760 DOMAIN CAN PRODUCE THIS. AND IT 621 00:29:56,760 --> 00:29:58,600 TURNS OUT ALL THE FAMILY MEMBERS 622 00:29:58,600 --> 00:30:02,680 FROM A-F IF YOU WILL. THIS IS 623 00:30:02,680 --> 00:30:05,920 AN F MEMBER. IT'S DOMAIN CAN 624 00:30:05,920 --> 00:30:12,080 ALL INDUCE POOR FORMATION AND 625 00:30:12,080 --> 00:30:13,960 PIE ROP TO SIS AND THEY CAN BE 626 00:30:13,960 --> 00:30:15,480 ACTIVATED BY DIFFERENT KINDS OF 627 00:30:15,480 --> 00:30:15,720 ENZYMES. 628 00:30:15,720 --> 00:30:21,400 I LIST A FEW ON THE SIDE OF THIS 629 00:30:21,400 --> 00:30:28,080 SLIDE. SO SO WHY PIE PROP SEW 630 00:30:28,080 --> 00:30:30,160 SIS. IT IS VERY DIFFERENT FROM 631 00:30:30,160 --> 00:30:33,440 APOP SEW SIS BECAUSE IT CAN 632 00:30:33,440 --> 00:30:37,240 DANGER THE MITOCHONDRIA AND 633 00:30:37,240 --> 00:30:39,400 THINGS CAN DO OUT AND IN. IT 634 00:30:39,400 --> 00:30:43,840 CAN GO OUT. HMGB 1. A LOT OF 635 00:30:43,840 --> 00:30:45,720 DIFFERENT DANGER MOLECULES CAN 636 00:30:45,720 --> 00:30:48,560 GO OUT SO THEY'RE VERY IMMUNO 637 00:30:48,560 --> 00:30:51,200 LOGICALLY HOT BY CONTRAST 638 00:30:51,200 --> 00:30:53,040 APOPTOSIS IS IMMUNO LOGICALLY 639 00:30:53,040 --> 00:30:56,080 COLD SO THEY DON'T REALLY ALERT 640 00:30:56,080 --> 00:30:59,480 THE IMMUNE SYSTEM TO ATTRACT THE 641 00:30:59,480 --> 00:31:02,760 ATTENTION THERE. SO OVER THE 642 00:31:02,760 --> 00:31:05,440 YEARS WE HAVE SOLVED TWO 643 00:31:05,440 --> 00:31:10,480 DIFFERENT GASDERMIN NT -- N 644 00:31:10,480 --> 00:31:12,920 TERMINAL DOMAIN POOR STRUCTURES. 645 00:31:12,920 --> 00:31:15,880 I JUST WANT TO DRAW YOUR 646 00:31:15,880 --> 00:31:19,880 ATTENTION THAT GASDERMIN PORES 647 00:31:19,880 --> 00:31:21,880 MAY BE FORMED IN TWO DIFFERENT 648 00:31:21,880 --> 00:31:23,880 STEPS. ONE IS A PREPORE. WHAT 649 00:31:23,880 --> 00:31:25,360 IS A PREPORE? THIS IS THE 650 00:31:25,360 --> 00:31:27,720 YELLOW HERE IS A PREPORE 651 00:31:27,720 --> 00:31:30,280 STRUCTURE. SO THE PREPORE IS 652 00:31:30,280 --> 00:31:32,160 ALSO ALREADY O LIG MA RISED ON 653 00:31:32,160 --> 00:31:33,560 THE MEMBRANE BUT IT HAS TO 654 00:31:33,560 --> 00:31:35,960 INSERT INTO THE MEMBRANE SO AS 655 00:31:35,960 --> 00:31:37,400 YOU CAN SEE HERE. 656 00:31:37,400 --> 00:31:40,800 THE INSERTED PART OF THE 657 00:31:40,800 --> 00:31:42,440 STRUCTURE IS A BETA BARREL. 658 00:31:42,440 --> 00:31:44,720 THIS IS A LARGE BETA BARREL THAT 659 00:31:44,720 --> 00:31:53,160 CONTAINS A LOT. 33 TIMES FOUR 660 00:31:53,160 --> 00:31:54,680 BETA STRANDS THAT MAKES THIS 661 00:31:54,680 --> 00:31:56,680 LARGE BETA BARREL SO THIS CAN BE 662 00:31:56,680 --> 00:31:59,560 FORMED AND THEN INSERTION CAN 663 00:31:59,560 --> 00:32:01,920 OCCUR IN ORDER TO FORM THE PORE 664 00:32:01,920 --> 00:32:03,360 STRUCTURE. THIS WILL BECOME 665 00:32:03,360 --> 00:32:05,080 IMPORTANT WHEN I TALK ABOUT THE 666 00:32:05,080 --> 00:32:07,200 REMAINING PART OF THE TALK. 667 00:32:07,200 --> 00:32:11,600 SOME OF IT UNPUBLISHED. SO THE 668 00:32:11,600 --> 00:32:14,360 SUBUNIT OF GASDERMIN LOOKS 669 00:32:14,360 --> 00:32:16,600 REALLY LIKE A LEFT HAND. THIS 670 00:32:16,600 --> 00:32:19,560 IS THE INACTIVE STRUCTURE LIKE 671 00:32:19,560 --> 00:32:21,840 THIS. THIS IS LIKE THE GLOBULAR 672 00:32:21,840 --> 00:32:24,480 DOMAIN AND DURING INSERTION YOU 673 00:32:24,480 --> 00:32:26,360 JUST HAVE YOUR FOUR FINGERS AS 674 00:32:26,360 --> 00:32:28,840 THE TWO -- PAIRS OF BETA HAIR 675 00:32:28,840 --> 00:32:32,240 PINS THAT GOES INTO THE 676 00:32:32,240 --> 00:32:34,560 MEMBRANE. ONCE YOU INSERT YOUR 677 00:32:34,560 --> 00:32:37,120 FINGERS LIKE THIS THE PORE IS 678 00:32:37,120 --> 00:32:40,240 FORMED AND THE PORE BECOMES THIS 679 00:32:40,240 --> 00:32:41,400 TRANSMEMBRANE PORE THAT CAN 680 00:32:41,400 --> 00:32:45,440 RELEASE THE CYTOKINE OR OTHER 681 00:32:45,440 --> 00:32:48,040 MOLECULES SO THIS IS A 682 00:32:48,040 --> 00:32:50,640 SIMULATION OF THE EVENTS IN THIS 683 00:32:50,640 --> 00:32:53,040 PREPORE TO PORE TRANSITION. 684 00:32:53,040 --> 00:32:55,440 THIS -- YELLOW STARTS WITH A 685 00:32:55,440 --> 00:32:57,400 PREPORE AND THEN THE STRANDS 686 00:32:57,400 --> 00:33:04,800 INSERT INTO A PORE. SO ONCE WE 687 00:33:04,800 --> 00:33:07,720 SOLVED THE PORE STRUCTURE THERE 688 00:33:07,720 --> 00:33:09,560 WAS ONE QUESTION THAT BUGGED US 689 00:33:09,560 --> 00:33:16,520 THAT IS THIS PARADOX. PROTO 690 00:33:16,520 --> 00:33:17,920 LIEU KIN ONE WHICH IS ALSO VERY 691 00:33:17,920 --> 00:33:22,920 SMALL CANNOT BE RELEASED FROM 692 00:33:22,920 --> 00:33:24,000 LIVE CELLS SO THESE LIVE CELL 693 00:33:24,000 --> 00:33:26,240 CONSIST STILL HAVE PORES ON THE 694 00:33:26,240 --> 00:33:29,560 SURFACE AND THEY CAN RELEASE 695 00:33:29,560 --> 00:33:33,400 MATURE ONE. WHICH IS 18 BUT 696 00:33:33,400 --> 00:33:35,560 THEY CANNOT RELEASE THE PROTO 697 00:33:35,560 --> 00:33:37,720 SAY TO SIGN AND THESE ARE THE 698 00:33:37,720 --> 00:33:39,800 DATA SHOWN HERE SO THE PRODOES 699 00:33:39,800 --> 00:33:41,360 NOT COME OUT BUT THE MATURE 700 00:33:41,360 --> 00:33:45,840 CYTOKINES COME OUT SIZE-WISE 701 00:33:45,840 --> 00:33:46,520 THEY ARE ALL ABOUT -- 702 00:33:46,520 --> 00:33:49,240 IN TERMS OF DIAMETER A QUARTER 703 00:33:49,240 --> 00:33:50,880 OR LESS OF THE BIG PORE 704 00:33:50,880 --> 00:33:52,320 STRUCTURE. LIKE THIS SIZE HERE 705 00:33:52,320 --> 00:33:54,200 FOR A BIG PORE. BUT IT CANNOT 706 00:33:54,200 --> 00:33:57,440 COME OUT. SO A PROMINENT 707 00:33:57,440 --> 00:34:01,400 FEATURE OF THE PROCYTOKINE 708 00:34:01,400 --> 00:34:03,880 THAT'S ABSENT IS THIS NEGATIVE 709 00:34:03,880 --> 00:34:06,560 CHARGE TO SO THE PROPEPTIDE HERE 710 00:34:06,560 --> 00:34:08,400 IS ACTUALLY EXTREMELY NEGATIVELY 711 00:34:08,400 --> 00:34:09,800 CHARGED SO WE THOUGHT IS IT 712 00:34:09,800 --> 00:34:13,360 POSSIBLE, THERE'S SOME KIND OF A 713 00:34:13,360 --> 00:34:15,560 CHARGE SELECTION FOR THE 714 00:34:15,560 --> 00:34:19,200 GASDERMIN PORE SELECTIVITY. SO 715 00:34:19,200 --> 00:34:23,360 GASDERMIN D IF YOU LOOK AT THE 716 00:34:23,360 --> 00:34:26,040 PI OF GASDERMIN IS ACTUALLY 8 717 00:34:26,040 --> 00:34:27,000 POINT SOMETHING SO YOU WOULD -- 718 00:34:27,000 --> 00:34:32,960 AT LEAST I DID NOT, I NEVER 719 00:34:32,960 --> 00:34:35,400 THOUGHT THIS COULD BE HAVE ANY 720 00:34:35,400 --> 00:34:38,400 NEGATIVE CHARGE IN THIS PROTEIN. 721 00:34:38,400 --> 00:34:39,720 IF YOU LOOK AT THE CHARGE 722 00:34:39,720 --> 00:34:42,840 DISTRIBUTION ON THE ENTIRE PORE, 723 00:34:42,840 --> 00:34:44,120 ALL THE STRONG POSITIVE CHARGES 724 00:34:44,120 --> 00:34:46,360 ARE FACING THE MEMBRANE AND 725 00:34:46,360 --> 00:34:48,680 THAT'S BECAUSE THEY'RE BINDING 726 00:34:48,680 --> 00:34:50,120 ACIDIC LIPIDS AND INSIDE THE 727 00:34:50,120 --> 00:34:52,200 PORE YOU SEE INSIDE THE PORE IS 728 00:34:52,200 --> 00:34:56,000 ACTUALLY VERY ACIDIC. AND WHEN 729 00:34:56,000 --> 00:34:59,200 YOU CALCULATE THE ELECTROSTATIC 730 00:34:59,200 --> 00:35:01,160 POTENTIAL OF THE WILD TYPE 731 00:35:01,160 --> 00:35:02,960 VERSUS MUTANT, IF YOU WANT 732 00:35:02,960 --> 00:35:06,320 ACIDIC PATCH ONE OR ACIDIC PATCH 733 00:35:06,320 --> 00:35:08,760 TWO AND THREE WHEN YOU CALCULATE 734 00:35:08,760 --> 00:35:09,920 THE ELECTROSTATIC POTENTIAL YOU 735 00:35:09,920 --> 00:35:14,880 DO SEE THAT WHEN YOU MUTATE. 736 00:35:14,880 --> 00:35:18,960 YOU HAVE REDUCED AMOUNT OF 737 00:35:18,960 --> 00:35:20,360 ACIDIC ELECTROSTATIC POTENTIAL 738 00:35:20,360 --> 00:35:23,760 SO WE DID A LOT OF DIFFERENT 739 00:35:23,760 --> 00:35:25,400 TESTS BUT I WANT TO SHOW YOU 740 00:35:25,400 --> 00:35:29,000 THIS LAST TEST WE DID IN CELLS 741 00:35:29,000 --> 00:35:33,040 IN WHICH WE USED THE CELLS AND 742 00:35:33,040 --> 00:35:33,400 RECON. 743 00:35:33,400 --> 00:35:34,480 >> THE STUDENT: CONSTITUTE WITH 744 00:35:34,480 --> 00:35:35,600 EITHER THE WILD TYPE, SORRY, I'M 745 00:35:35,600 --> 00:35:38,480 SO TERRIBLE WITH DRAWING THERE. 746 00:35:38,480 --> 00:35:42,000 WILD TYPE OR THE ACIDIC PATCH 747 00:35:42,000 --> 00:35:44,920 MUTANT OF GASDERMIN. AND THEN 748 00:35:44,920 --> 00:35:47,320 STIMULATE THE CELLS WITH VERY 749 00:35:47,320 --> 00:35:49,400 LOW DOSE NIGER RICIN FOR A LONG 750 00:35:49,400 --> 00:35:51,360 TIME WHICH THE CELLS DO NOT DIE 751 00:35:51,360 --> 00:35:53,800 AND LOOK AT THE RELEASE OF 752 00:35:53,800 --> 00:35:57,400 INTERLIEU ONE ONE VERSUS THE 753 00:35:57,400 --> 00:35:58,080 PROTEIN INTERLEUKIN ONE SO AS 754 00:35:58,080 --> 00:36:00,520 YOU CAN SEE HERE WITH THE WILD 755 00:36:00,520 --> 00:36:03,360 TYPE RECONSTITUTION YOU HAVE 756 00:36:03,360 --> 00:36:05,120 ONLY THE MATURE CYTOKINE COMING 757 00:36:05,120 --> 00:36:06,560 OUT VERY NICELY. 758 00:36:06,560 --> 00:36:09,400 WHEREAS FOR THE CELLS THAT'S 759 00:36:09,400 --> 00:36:10,880 RECONSTITUTED WITH THIS ACIDIC 760 00:36:10,880 --> 00:36:16,160 PATCH MUTANT YOU SEE 761 00:36:16,160 --> 00:36:17,160 PROINTERLEUKIN ONE COMING OUT 762 00:36:17,160 --> 00:36:19,000 AND YOU ONLY HAVE A LITTLE BIT 763 00:36:19,000 --> 00:36:21,880 OF THE MATURE INTERLEUKIN ONE 764 00:36:21,880 --> 00:36:25,120 AND THAT'S BECAUSE THE 765 00:36:25,120 --> 00:36:25,760 PROCYTOKINES ARE ALREADY OUT. 766 00:36:25,760 --> 00:36:27,160 SO THEY DON'T HAVE A CHANCE TO 767 00:36:27,160 --> 00:36:30,320 BE PROCESSED. REALLY SUGGESTING 768 00:36:30,320 --> 00:36:33,200 THAT THIS SELECTIVITY FOR THE 769 00:36:33,200 --> 00:36:35,560 RELEASE OF MATURE CYTOKINE AND 770 00:36:35,560 --> 00:36:37,160 PERHAPS ALSO A LOT OF OTHER 771 00:36:37,160 --> 00:36:38,640 THINGS IS GOVERNED BY THIS 772 00:36:38,640 --> 00:36:41,400 CHARGE DISTRIBUTION WHAT WE CALL 773 00:36:41,400 --> 00:36:45,400 ELECTROSTATIC FILTERING IN THE 774 00:36:45,400 --> 00:36:50,480 GASDERMIN CARGO RELEASE. YEAH, 775 00:36:50,480 --> 00:36:52,240 WHY DOES THE PORE HAVE THE 776 00:36:52,240 --> 00:36:55,960 SELECTIVITY IS IT REALLY EVOLVED 777 00:36:55,960 --> 00:36:57,440 ONLY FOR INTERLEUKIN ONE RELEASE 778 00:36:57,440 --> 00:37:00,200 OR ALSO IMPORTANT FOR SOME OTHER 779 00:37:00,200 --> 00:37:04,080 ASPECT OF THE BIOLOGY REMAINS 780 00:37:04,080 --> 00:37:07,400 UNCLEAR. BUT ONE DISCUSSION 781 00:37:07,400 --> 00:37:15,160 THAT I HAD I THINK WI. HE SAID 782 00:37:15,160 --> 00:37:17,400 LOT OF DANGER MOLECULES ARE 783 00:37:17,400 --> 00:37:18,880 POSITIVELY CHARGED WHICH MEANS 784 00:37:18,880 --> 00:37:20,480 THEY CAN BE RELEASED FROM THE 785 00:37:20,480 --> 00:37:23,640 PORE. SO WE SHALL SEE IF THAT 786 00:37:23,640 --> 00:37:25,400 IS THE CASE WHEN WE PERFORM THE 787 00:37:25,400 --> 00:37:27,360 EXPERIMENT WHICH IS ONGOING. 788 00:37:27,360 --> 00:37:28,760 SO, I AM JUST LOOKING AT THE 789 00:37:28,760 --> 00:37:30,800 TIME. OKAY, SO IN THE LAST 790 00:37:30,800 --> 00:37:35,640 PART, I WANT TO TELL YOU AN 791 00:37:35,640 --> 00:37:37,400 UNPUBLISHED STORY. ON THIS 792 00:37:37,400 --> 00:37:39,480 QUESTION THAT HAS BUGGED US FOR 793 00:37:39,480 --> 00:37:41,400 AWHILE. SO WE KNOW BACK IN 2016 794 00:37:41,400 --> 00:37:44,800 WHEN WE DISCOVERED THAT 795 00:37:44,800 --> 00:37:48,120 GASDERMIN CAN FORM PORES WE 796 00:37:48,120 --> 00:37:50,840 MUTATED OTHER CYSTEINE RESIDUES 797 00:37:50,840 --> 00:37:54,160 AND FOUND THAT C191A IS 798 00:37:54,160 --> 00:37:56,960 DEFECTIVE. IF YOU TRANSFECT 799 00:37:56,960 --> 00:37:58,760 THIS MUTANT YOU CANNOT KILL 800 00:37:58,760 --> 00:38:01,400 CELLS BUT WE DIDN'T UNDERSTAND 801 00:38:01,400 --> 00:38:03,120 WHY AND EVEN AFTER WE HAD THIS 802 00:38:03,120 --> 00:38:04,880 STRUCTURE WE KNOW THIS RESIDUE 803 00:38:04,880 --> 00:38:07,480 IS AT THE TIP OF THE BETA HAIR 804 00:38:07,480 --> 00:38:10,320 PIN. RIGHT AT THE TIP BUT 805 00:38:10,320 --> 00:38:12,840 THERE'S NO DULL SULFIDE AND THE 806 00:38:12,840 --> 00:38:14,640 RESIDUE IS VERY FAR APART FROM 807 00:38:14,640 --> 00:38:15,560 EACH OTHER IN THE PORE. 808 00:38:15,560 --> 00:38:17,400 WE DON'T KNOW WHAT IS THE 809 00:38:17,400 --> 00:38:18,400 FUNCTION OF THIS THING. SO 810 00:38:18,400 --> 00:38:20,760 GIVEN THAT THIS IS A MEMBRANE 811 00:38:20,760 --> 00:38:22,760 PROTEIN I ACTUALLY SUSPECTED 812 00:38:22,760 --> 00:38:26,400 MAYBE IT IS LIP DATED BUT WE 813 00:38:26,400 --> 00:38:28,560 REALLY DIDN'T DO AFTER IT UNTIL 814 00:38:28,560 --> 00:38:31,480 A BRAVE POSTDOC DECIDES TO TRY 815 00:38:31,480 --> 00:38:32,160 THAT EXPERIMENTALLY. 816 00:38:32,160 --> 00:38:35,320 SO INDEED, TA TURNED OUT TO BE 817 00:38:35,320 --> 00:38:39,840 THE CASE. THE CYSTEINE RESIDUE 818 00:38:39,840 --> 00:38:42,560 IS POE MODULATED. THIS IS ONE 819 00:38:42,560 --> 00:38:45,560 OF THE LIP DAGS FORM BUT POE 820 00:38:45,560 --> 00:38:46,760 MODULATION IS I BELIEVE I 821 00:38:46,760 --> 00:38:50,000 BELIEVE THE ONLY LIPID 822 00:38:50,000 --> 00:38:53,400 MODIFICATION THAT IS REVERSIBLE 823 00:38:53,400 --> 00:38:56,760 CAN BE REGULATED. SO WE USE A 824 00:38:56,760 --> 00:38:59,960 NUMBER OF METHODS TO LOOK AT 825 00:38:59,960 --> 00:39:06,760 THIS -- PAL MY TO MODIFICATION. 826 00:39:06,760 --> 00:39:09,400 ONE IS CALLED ACYL BIOTIN 827 00:39:09,400 --> 00:39:11,080 EXCHANGE WHICH BASICALLY YOU 828 00:39:11,080 --> 00:39:13,440 MODIFY ALL THE FREE CYSTEINES IN 829 00:39:13,440 --> 00:39:21,800 THE CELL. THEN YOU CLEAVE THIS 830 00:39:21,800 --> 00:39:25,400 BOUND -- SO PAL MY TOLATION IS 831 00:39:25,400 --> 00:39:29,200 WITH THE EXISTING RESIDUES USING 832 00:39:29,200 --> 00:39:32,240 THIS HYDROXYL AMINE AND THEN 833 00:39:32,240 --> 00:39:35,640 AFTER YOU TREAT THIS -- THESE 834 00:39:35,640 --> 00:39:39,320 CELLS WITH HYDROXYL AMINE YOU 835 00:39:39,320 --> 00:39:41,280 LEAVE THE THE GROUP WITH THE 836 00:39:41,280 --> 00:39:43,760 FILE AND YOU CAN PUT IN ANYTHING 837 00:39:43,760 --> 00:39:45,400 THAT YOU WANT A BIOTIN THAT HAS 838 00:39:45,400 --> 00:39:47,120 A FREE FILE AND YOU CAN PULL IT 839 00:39:47,120 --> 00:39:52,800 DOWN. SO AS YOU CAN SEE IT IS 840 00:39:52,800 --> 00:39:55,320 MALL MY TOLATED BUT NOT C 841 00:39:55,320 --> 00:39:57,680 TERMINUS. IT IS NOT PAL MY 842 00:39:57,680 --> 00:39:59,760 TOLATED. OTHER GASDERMINS ARE 843 00:39:59,760 --> 00:40:02,040 ALSO PAL MY TOLATED SO WE START 844 00:40:02,040 --> 00:40:05,320 TO THINK THAT MAYBE THIS IS A 845 00:40:05,320 --> 00:40:08,080 GENERAL PHENOMENA FOR THE ENTIRE 846 00:40:08,080 --> 00:40:11,080 GASDERMIN FAMILY SO 847 00:40:11,080 --> 00:40:18,520 INTERESTINGLY, IF YOU TAKE A 848 00:40:18,520 --> 00:40:21,280 MACROPHAGE, YOU PRIME THE CELL 849 00:40:21,280 --> 00:40:23,480 WITH LIE POE POLY SACK RAH, LPS, 850 00:40:23,480 --> 00:40:25,760 YOU SEE A LITTLE BIT OF INCREASE 851 00:40:25,760 --> 00:40:27,880 IN PAL MY TOLATION. WITHOUT IT 852 00:40:27,880 --> 00:40:30,840 YOU SEE NOTHING, NOTHING CAN BE 853 00:40:30,840 --> 00:40:33,000 DETECTED AND THEN WHEN YOU TREAT 854 00:40:33,000 --> 00:40:40,680 A CELL WITH LPS PLUS NEM YOU SEE 855 00:40:40,680 --> 00:40:43,480 A GREAT INCREASE. BOTH ON THE 856 00:40:43,480 --> 00:40:46,720 CELLS AND THE N TERMINUS WHEREAS 857 00:40:46,720 --> 00:40:48,520 AT LEAST UNDER THIS CONDITION 858 00:40:48,520 --> 00:40:51,240 THERE'S NO GAME IN PROCESSING 859 00:40:51,240 --> 00:40:52,720 SUGGESTING THAT THE ENTIRE 860 00:40:52,720 --> 00:40:54,520 UPSTREAM EVENTS WERE NOT 861 00:40:54,520 --> 00:40:56,000 AFFECTED. ONLY THE DOWNSTREAM 862 00:40:56,000 --> 00:40:59,240 EVENTS ON THE LIPID MODIFICATION 863 00:40:59,240 --> 00:41:01,400 OF GASDERMIN M.D. SO WE MAPPED 864 00:41:01,400 --> 00:41:03,560 WHICH RESIDUE OF COURSE YOU KNOW 865 00:41:03,560 --> 00:41:10,440 THE ANSWER ALREADY IS C191. I 866 00:41:10,440 --> 00:41:12,480 COME BACK TO THAT BUT IT TURNED 867 00:41:12,480 --> 00:41:14,080 OUT TO BE AN INTERESTING 868 00:41:14,080 --> 00:41:14,960 RESIDUE. IT'S INVOLVED IN SO 869 00:41:14,960 --> 00:41:18,720 MANY MYSTERIES ABOUT THIS 870 00:41:18,720 --> 00:41:21,400 PROTEIN. SO WE THEN VALIDATE 871 00:41:21,400 --> 00:41:24,280 THIS IN VITRO TO SEE WHETHER THE 872 00:41:24,280 --> 00:41:26,160 PAL MY TOLATED GASDERMIN M.D. IS 873 00:41:26,160 --> 00:41:29,720 MORE ACTIVATED. THIS IS A 874 00:41:29,720 --> 00:41:32,320 LYSOSOME USING PURIFIED 875 00:41:32,320 --> 00:41:33,320 PROTEINS. IF YOU PURIFY THESE 876 00:41:33,320 --> 00:41:34,880 PROTEINS FROM BACTERIA, I 877 00:41:34,880 --> 00:41:37,400 BELIEVE THE ORANGE HERE IS THE 878 00:41:37,400 --> 00:41:39,440 WILD TYPE FROM BACTERIA. THE 879 00:41:39,440 --> 00:41:42,640 GREEN IS THE C191 MUTANT FROM 880 00:41:42,640 --> 00:41:44,920 BACTERIA. IN BACTERIA THAT 881 00:41:44,920 --> 00:41:46,880 MEANS THEY'RE NOT PAL MY TO 882 00:41:46,880 --> 00:41:49,080 LATED. YOU CAN SEE THIS IS THE 883 00:41:49,080 --> 00:41:52,280 RITE OF LIPOSOME HAECAGE OF THE 884 00:41:52,280 --> 00:41:54,400 RATE THAT THESE PROTEINS DPAJ 885 00:41:54,400 --> 00:41:57,400 THE LIPOSOME TO RELEASE THE DYE. 886 00:41:57,400 --> 00:42:01,080 BUT IF YOU USE MAMMAL YAN 887 00:42:01,080 --> 00:42:03,760 GASDERMIN M.D. THIS IS ABOUT 50% 888 00:42:03,760 --> 00:42:06,680 PAL MY TOLATED. ABOUT 50%. YOU 889 00:42:06,680 --> 00:42:08,680 CAN ALREADY SEE ABOUT 10 FOLD 890 00:42:08,680 --> 00:42:11,680 INCREASE IN THE RATE OF PAL MY 891 00:42:11,680 --> 00:42:15,560 TOLATION SUGGESTING INDEED THIS 892 00:42:15,560 --> 00:42:17,400 GROUP REALLY HELPED TO INSERT 893 00:42:17,400 --> 00:42:20,520 GASDERMIN M.D. INTO THE MEMBRANE 894 00:42:20,520 --> 00:42:23,680 SO FUNCTIONALLY, LIKE, THE SAME 895 00:42:23,680 --> 00:42:26,520 THING, THE C191 A MUTANT IS 896 00:42:26,520 --> 00:42:28,560 DEFECTIVE AND WHEN WE LOOKED AT 897 00:42:28,560 --> 00:42:33,400 THEM BY CELLULAR IMAGING, WILD 898 00:42:33,400 --> 00:42:36,760 TYPE, CAN GO TO THE CELL 899 00:42:36,760 --> 00:42:39,680 SURFACE. YOU CAN SEE FROM THESE 900 00:42:39,680 --> 00:42:42,200 STRUCTURES THAT'S ON THE 901 00:42:42,200 --> 00:42:44,480 MEMBRANE, WHEREAS THE MUTANT, 902 00:42:44,480 --> 00:42:47,600 THE C1 AND 191A MUTANT IS NOT 903 00:42:47,600 --> 00:42:50,200 REALLY MEMBRANE LOCALIZED AT ALL 904 00:42:50,200 --> 00:42:52,560 SO IT'S EFFECTING LOCALIZATION 905 00:42:52,560 --> 00:42:56,000 AND PERHAPS ALSO PORE FORMATION. 906 00:42:56,000 --> 00:42:57,400 SO THIS IS -- THIS I THINK IS 907 00:42:57,400 --> 00:42:59,720 WHERE IT GETS REALLY 908 00:42:59,720 --> 00:43:03,080 INTERESTING. SO BECAUSE THIS IS 909 00:43:03,080 --> 00:43:05,200 ESTER BOUND SO WE WORKED WITH 910 00:43:05,200 --> 00:43:06,480 OUR ESTER BOUND BEFORE AND WE 911 00:43:06,480 --> 00:43:09,280 KNOW THAT ANY REDUCING AGENT, 912 00:43:09,280 --> 00:43:11,360 ANYTHING THAT HAS A FREE FILE 913 00:43:11,360 --> 00:43:15,040 CAN ATTACK THIS. AND DESTROY 914 00:43:15,040 --> 00:43:17,680 THIS FILE ESTER. SO WE THOUGHT, 915 00:43:17,680 --> 00:43:21,440 HUH, THIS PROBABLY MEANS THAT 916 00:43:21,440 --> 00:43:24,960 THIS IS SENSITIVE TO THE REDOC 917 00:43:24,960 --> 00:43:27,560 STATE OF A CELL. IF YOU HAVE 918 00:43:27,560 --> 00:43:28,480 THIS IT WILL NOT STAY IF YOU 919 00:43:28,480 --> 00:43:30,480 HAVE A MORE OXIDIZING 920 00:43:30,480 --> 00:43:31,200 ENVIRONMENT MAYBE THIS WILL STAY 921 00:43:31,200 --> 00:43:32,920 AND STAY LONGER SO THEN WE 922 00:43:32,920 --> 00:43:35,680 THOUGHT, OKAY, MAYBE THEN THIS 923 00:43:35,680 --> 00:43:38,120 WILL LINK IT TO THE GENERATION 924 00:43:38,120 --> 00:43:41,400 OF ROS WITH CHANGES, THE REDOCK 925 00:43:41,400 --> 00:43:44,960 STATE OF THE CELL SO WHAT WE DID 926 00:43:44,960 --> 00:43:48,440 THEN WAS TO USE THE SAME TH 927 00:43:48,440 --> 00:43:51,720 CELLS BUT ALSO INSTEAD OF USING 928 00:43:51,720 --> 00:43:54,120 JUST LPS PLUS NIGER RICIN WE 929 00:43:54,120 --> 00:43:58,600 ALSO TREATED CELLS WITH ROW TE 930 00:43:58,600 --> 00:44:02,800 KNOWN IS ONE. ANTIMY SEEN A. 931 00:44:02,800 --> 00:44:06,800 THESE TWO ARE INHIBITORS OF THE 932 00:44:06,800 --> 00:44:08,560 MITOCHONDRIA RESPIRATORY 933 00:44:08,560 --> 00:44:09,280 COMPLEX. ONE VERSUS THREE AND 934 00:44:09,280 --> 00:44:13,920 THEY ALL INDUCE ROS IN THE 935 00:44:13,920 --> 00:44:16,880 MITOCHONDRIA SO THESE TWO AS YOU 936 00:44:16,880 --> 00:44:20,440 CAN SEE HERE INCREASES PAL MY 937 00:44:20,440 --> 00:44:21,880 TOLATION BUT NO CHANGE IN THE 938 00:44:21,880 --> 00:44:23,720 CLEAVAGE. IF YOU TREAT THE 939 00:44:23,720 --> 00:44:27,560 CELLS IN CONTRAST USING ACETYL 940 00:44:27,560 --> 00:44:28,440 CYSTEINE, THIS IS A REDUCING 941 00:44:28,440 --> 00:44:31,640 AGENT YOU REDUCE THE AMOUNT OF 942 00:44:31,640 --> 00:44:33,440 PAL MY TOLATION AMOUNT. AND 943 00:44:33,440 --> 00:44:36,480 THIS IS ANOTHER INHIBITOR THAT 944 00:44:36,480 --> 00:44:42,920 ALSO REDUCES THE CELLULAR ROS. 945 00:44:42,920 --> 00:44:44,320 SO THIS IS ALREADY SAYING, OKAY, 946 00:44:44,320 --> 00:44:48,280 IF YOU HAVE AN ROS ENHANCER YOU 947 00:44:48,280 --> 00:44:51,640 HAVE MORE PAL MY TO LAGS. IF 948 00:44:51,640 --> 00:44:53,320 YOU USE AN EXHIBIT TOR YOU HAVE 949 00:44:53,320 --> 00:44:55,200 LESS. WHAT HAPPENED TO THE CELL 950 00:44:55,200 --> 00:45:00,240 UNDER THOSE CAN BES, HAD THE ROK 951 00:45:00,240 --> 00:45:01,880 REALLY INCREASED OR DECREASED. 952 00:45:01,880 --> 00:45:03,840 THIS IS WHERE YOU TREAT THE 953 00:45:03,840 --> 00:45:06,360 CELLS. THIS IS BY IMAGING THE 954 00:45:06,360 --> 00:45:09,160 AMOUNT OF DWELLULAR ROS THAT YOU 955 00:45:09,160 --> 00:45:11,680 OBSERVE OR THE MITOCHONDRIA OR 956 00:45:11,680 --> 00:45:15,920 AS YOU SOEB. IF YOU TREAT A 957 00:45:15,920 --> 00:45:22,720 CELL ALSO WITH THIS TWO YOU 958 00:45:22,720 --> 00:45:24,920 REDUCE THE ROS. AND IF YOU 959 00:45:24,920 --> 00:45:28,560 TREAT THE CELLS WITH THE ONE 960 00:45:28,560 --> 00:45:32,240 THAT INCREASE PAL MY TOLATION 961 00:45:32,240 --> 00:45:40,120 YOU ALSO GREATLY INKRECREASE TH 962 00:45:40,120 --> 00:45:41,720 ROS SUGGESTING THEY'RE COUPLED. 963 00:45:41,720 --> 00:45:45,280 ONE ENHANCING THE OTHER. SO 964 00:45:45,280 --> 00:45:50,680 THEN WE LOOKED AT AMOUNT OF 23 965 00:45:50,680 --> 00:45:53,000 HUMAN TRANSFERASES AND THEY'RE 966 00:45:53,000 --> 00:45:56,760 ALL NAMED THIS GUY HERE BECAUSE 967 00:45:56,760 --> 00:45:59,240 THEY HAVE A 2H AND THE CYSTEINE 968 00:45:59,240 --> 00:46:01,520 AT THE ACTIVE SITE. WE TESTED 969 00:46:01,520 --> 00:46:05,400 ALL OF THEM AND FOUND SEVERAL OF 970 00:46:05,400 --> 00:46:11,720 THEM MAY ENHANCE THE PAL MY 971 00:46:11,720 --> 00:46:14,200 TOLATION OF THIS. AND JUDGING 972 00:46:14,200 --> 00:46:17,160 FROM THE -- SIT PROTEIN ATLAS? 973 00:46:17,160 --> 00:46:21,920 WEST SIDE? WE CHOSE TO 974 00:46:21,920 --> 00:46:22,840 INVESTIGATE FURTHER THESE FIVE 975 00:46:22,840 --> 00:46:27,600 DIFFERENT PAL MY TO TILL 976 00:46:27,600 --> 00:46:29,400 TRANSFERASES SO WHEN WE KNOCKED 977 00:46:29,400 --> 00:46:33,400 DOWN ONE BY ONE THESE DIFFERENT 978 00:46:33,400 --> 00:46:37,200 PAL MITO TRANSFERASES. YOU SEE 979 00:46:37,200 --> 00:46:38,040 THIS IS IMPORTANT FOR THIS 980 00:46:38,040 --> 00:46:39,360 TRANSFORMATION AND NINE IS 981 00:46:39,360 --> 00:46:41,400 IMPORTANT AS WELL. SIMILARLY, 982 00:46:41,400 --> 00:46:45,440 THE KNOCK DOWN OF THESE 983 00:46:45,440 --> 00:46:48,880 TRANSFERASES ALSO REDUCE CELL 984 00:46:48,880 --> 00:46:52,000 DEATH, MEMBRANE LOCALIZATION AND 985 00:46:52,000 --> 00:46:56,760 EVERYTHING ELSE. SO THIS LED TO 986 00:46:56,760 --> 00:47:00,800 OUR MODEL ABOUT WHAT ROS AND PAL 987 00:47:00,800 --> 00:47:02,920 MY TOLATION MAY TOGETHER 988 00:47:02,920 --> 00:47:07,280 REGULATE GASDERMIN D ACTIVATION. 989 00:47:07,280 --> 00:47:14,560 SO SO THESE HAVE ALL BEEN SHOWN 990 00:47:14,560 --> 00:47:17,000 TO LOCALIZE AROUND THE GOLGI AND 991 00:47:17,000 --> 00:47:19,480 MAYBE ALSO THE ER AND MAYBE ALSO 992 00:47:19,480 --> 00:47:22,520 OTHER LOCATIONS SO WE THINK THAT 993 00:47:22,520 --> 00:47:28,120 WHEN GASDERMINS D IS NOT CLEAVED 994 00:47:28,120 --> 00:47:32,080 MAYBE THE MAIN SOURCE OF PAL MY 995 00:47:32,080 --> 00:47:33,320 TOLATION IS THROUGH THESE 996 00:47:33,320 --> 00:47:35,480 CYTOSOLIC ONES AND WHEN YOU HAVE 997 00:47:35,480 --> 00:47:38,040 HIGHER ROS YOU GET ADMINISTER. 998 00:47:38,040 --> 00:47:41,240 SO THIS LITTLE SKIG L HERE IS 999 00:47:41,240 --> 00:47:47,320 THE LIPID. AND THEN GASDERMIN D 1000 00:47:47,320 --> 00:47:52,400 CAN BE ACTIVATED OR CLEAVED BY 1001 00:47:52,400 --> 00:47:57,000 MATURE CAS CASE- 1 OR OTHER 1002 00:47:57,000 --> 00:48:02,840 CASPASES AND IF THE CLEAVED 1003 00:48:02,840 --> 00:48:06,120 GASDERMIN M.D. IS ALREADY PAL MY 1004 00:48:06,120 --> 00:48:07,760 TOLATED. IT WILL FORM A PREPORE 1005 00:48:07,760 --> 00:48:10,240 AND IMMEDIATELY TRANSIT INTO THE 1006 00:48:10,240 --> 00:48:14,000 PORE ITSELF. SO WHY DO I SAY 1007 00:48:14,000 --> 00:48:18,760 THAT? IF YOU LOOK AT THE 1008 00:48:18,760 --> 00:48:20,240 PREPORE AND WHERE THE 1009 00:48:20,240 --> 00:48:21,440 CALCULATION IS RIGHT AT THE TIP 1010 00:48:21,440 --> 00:48:24,040 OF THE INCERTIFICATED BITS THE 1011 00:48:24,040 --> 00:48:26,720 LIPID THAT IS -- THE LIPT THAT'S 1012 00:48:26,720 --> 00:48:29,440 ATTACHED TO THIS FINGERTIP HERE 1013 00:48:29,440 --> 00:48:31,560 I IMAGINE, AT LEAST, MAY HELP TO 1014 00:48:31,560 --> 00:48:34,520 DRAG THE INSERTION PROCESS SO 1015 00:48:34,520 --> 00:48:38,120 THAT EVERYTHING WILL GET INS 1016 00:48:38,120 --> 00:48:40,920 INSERTED INTO THE PORE. ANOTHER 1017 00:48:40,920 --> 00:48:44,600 REASON I SAY THIS IS JOHN'S 1018 00:48:44,600 --> 00:48:47,600 GROUP PUBLISHED A PAPER JUST 1019 00:48:47,600 --> 00:48:50,640 MAYBE ABOUT A YEAR AGO AND THEY 1020 00:48:50,640 --> 00:48:54,920 SHOWED THAT THIS MTOR RELATED 1021 00:48:54,920 --> 00:48:57,360 ROS IS IMPORTANT FOR THE 1022 00:48:57,360 --> 00:48:59,680 INSERTION OF THE PORE BUT NOT 1023 00:48:59,680 --> 00:49:02,760 FOR THE LOCALIZATION PER SE. SO 1024 00:49:02,760 --> 00:49:04,920 THIS I THINK, AT LEAST 1025 00:49:04,920 --> 00:49:09,440 MECHANISTICALLY CAN EXPLAIN 1026 00:49:09,440 --> 00:49:12,320 THAT. ROS WHICH EFFECTS SOME 1027 00:49:12,320 --> 00:49:13,600 MEMBRANE LOCALIZATION AND THE 1028 00:49:13,600 --> 00:49:15,320 FORMATION OF THE PREPORE BUT 1029 00:49:15,320 --> 00:49:16,200 MAYBE THE MOST IMPORTANT EFFECT 1030 00:49:16,200 --> 00:49:19,400 OF THIS IS TO DRAG THE PREPORE 1031 00:49:19,400 --> 00:49:23,720 INTO THE PORE TO FORM THIS 1032 00:49:23,720 --> 00:49:27,240 FUNCTIONING PORE. SO ONCE IT'S 1033 00:49:27,240 --> 00:49:28,880 CLEAVED IF THE PREPORE CAN ALSO 1034 00:49:28,880 --> 00:49:32,040 BE FORMED ON THE CELL MEMBRANE 1035 00:49:32,040 --> 00:49:34,280 THEN THE CELL MEMBRANE FORM OF 1036 00:49:34,280 --> 00:49:36,440 PAL MY TOLE TRANSFERASE, 1037 00:49:36,440 --> 00:49:39,360 ACTUALLY ONLY THE FIVE. OF ALL 1038 00:49:39,360 --> 00:49:40,680 THE TRANSFERASES FIVE IS THE 1039 00:49:40,680 --> 00:49:42,800 ONLY ONE KNOWN TO BE ON THE CELL 1040 00:49:42,800 --> 00:49:44,400 SURFACE AS WELL. I IMAGINE THE 1041 00:49:44,400 --> 00:49:46,040 FIVE WOULD BE THE MAJOR ONE THAT 1042 00:49:46,040 --> 00:49:49,080 WOULD CHANGE THE PREPORE TO A 1043 00:49:49,080 --> 00:49:52,640 PAL MODULATED PREPORE AND 1044 00:49:52,640 --> 00:49:55,120 FACILITATE ITS INSERTION. I'M 1045 00:49:55,120 --> 00:50:03,120 JUST LOOK AT MY TIME. SO IN 1046 00:50:03,120 --> 00:50:05,440 THEORY IT HAS BEEN SHOWN THAT 1047 00:50:05,440 --> 00:50:07,080 PAL MY TOLATED STUFF, THEY TEND 1048 00:50:07,080 --> 00:50:10,080 TO CLUSTER AND THIS IS BECAUSE 1049 00:50:10,080 --> 00:50:17,400 THESE LIPIDS, PALMITATE IS ALLEY 1050 00:50:17,400 --> 00:50:18,640 FATTIC. THEY ARE DOUBLE BOUND. 1051 00:50:18,640 --> 00:50:23,640 SO THEY'RE VERY ORDERED. THEY 1052 00:50:23,640 --> 00:50:25,000 CAN CAUSE PHASE SEPARATION IN 1053 00:50:25,000 --> 00:50:27,040 THE MEMBRANE SO THAT THE PORES 1054 00:50:27,040 --> 00:50:29,640 OR THE PAL MY TOLATED GROUPS 1055 00:50:29,640 --> 00:50:31,480 KNEW BECAMEMITOYLATED GROUPS KNW 1056 00:50:31,480 --> 00:50:33,440 BECAME PHASE SEPARATE INTO A 1057 00:50:33,440 --> 00:50:35,360 CLUSTERED ENTITY WITH MULTIPLE 1058 00:50:35,360 --> 00:50:37,440 PORES ALTOGETHER AND THIS IS 1059 00:50:37,440 --> 00:50:40,240 INDEED WHAT WE SAW ON THE LIGHT 1060 00:50:40,240 --> 00:50:41,720 IMAGING DATA. WE ACTUALLY DON'T 1061 00:50:41,720 --> 00:50:45,320 SEE A -- EVEN DISTRIBUTION OF 1062 00:50:45,320 --> 00:50:49,440 GASDERMIN ON THE CELL SUR 1063 00:50:49,440 --> 00:50:51,720 SURFACE. STRUCTURES. 1064 00:50:51,720 --> 00:50:53,120 WE WILL PROBABLY SEE A 1065 00:50:53,120 --> 00:50:57,400 CLUSTERING OF THE PORES. SO 1066 00:50:57,400 --> 00:51:01,200 INTERESTINGLY, A FEW YEARS AGO 1067 00:51:01,200 --> 00:51:03,080 WE PUBLISHED THIS DISULFUR RAM 1068 00:51:03,080 --> 00:51:05,360 WHICH IS A MODIFIER. GUESS WHAT 1069 00:51:05,360 --> 00:51:11,440 THEY ALL MODIFY. AND THE GROUP 1070 00:51:11,440 --> 00:51:12,960 AND KATE'S GROUP ALSO SHOWED 1071 00:51:12,960 --> 00:51:15,640 THESE OTHER MOLECULES AND THEY 1072 00:51:15,640 --> 00:51:19,840 ALL MODIFY CYSTEINE 191 AND THEY 1073 00:51:19,840 --> 00:51:26,520 ALL INHIBIT GASDERMIN M.D. AND 1074 00:51:26,520 --> 00:51:30,400 WE FIGURE THEY INHIBIT THE CELLS 1075 00:51:30,400 --> 00:51:33,600 BY COMPETING WITH PAL MY 1076 00:51:33,600 --> 00:51:37,400 TOLATION AND DIMETHYL FUME MA 1077 00:51:37,400 --> 00:51:42,680 RATE WORKS AND FUMARATE WORKS A 1078 00:51:42,680 --> 00:51:46,400 MOLECULE THAT GETS UPREGULATED 1079 00:51:46,400 --> 00:51:47,280 DURING REGULAR DAGS. 1080 00:51:47,280 --> 00:51:49,160 THIS MAY BE A NEGATIVE FEEDBACK 1081 00:51:49,160 --> 00:51:53,440 WITHIN THE CELL TO SUPPRESS 1082 00:51:53,440 --> 00:51:55,240 FURTHER INFLAMMASOME ACTIVATION 1083 00:51:55,240 --> 00:52:01,560 AFTER THE OCCURRENCE OF 1084 00:52:01,560 --> 00:52:04,040 INFLAMMASOME ACTIVATION. AS A 1085 00:52:04,040 --> 00:52:06,800 SUMMARY, I SHOWED YOU SOME DATA 1086 00:52:06,800 --> 00:52:12,320 NOW IN OUR P 3 PROJECT. 1087 00:52:12,320 --> 00:52:15,040 SUGGESTING THAT NLRP 3 IS 1088 00:52:15,040 --> 00:52:17,440 ACTIVATED THROUGH ONLY LARGE 1089 00:52:17,440 --> 00:52:21,440 CONFIRMATIONAL CHANGES THAT'S 1090 00:52:21,440 --> 00:52:22,600 AASSOCIATED WITH BINDING BUT 1091 00:52:22,600 --> 00:52:24,240 ALSO WITH TRAFFICKING AND 1092 00:52:24,240 --> 00:52:26,120 PERHAPS THESE STEPS CAN BE 1093 00:52:26,120 --> 00:52:27,040 TARGETED BY THERAPEUTICS. 1094 00:52:27,040 --> 00:52:30,840 AT THE MOMENT, ALL THE NLRP 3 1095 00:52:30,840 --> 00:52:33,400 DRUGS ARE TARGETING THE ADP 1096 00:52:33,400 --> 00:52:33,640 FORM. 1097 00:52:33,640 --> 00:52:37,400 TO KEEP IT IN THE ADP FORM SO IT 1098 00:52:37,400 --> 00:52:41,400 DOESN'T GET ACTIVATED AND THEN 1099 00:52:41,400 --> 00:52:44,320 WE SHOWED GASDERMIN PORE, EVEN 1100 00:52:44,320 --> 00:52:45,200 THOUGH IT'S SO LARGE WE WERE 1101 00:52:45,200 --> 00:52:49,120 PRETTY SURPRISED IT HAS SOME 1102 00:52:49,120 --> 00:52:50,760 SELECTIVITY. DESPITE ITS SIZE 1103 00:52:50,760 --> 00:52:51,880 IT DOES HAVE CERTAIN SELECTIVITY 1104 00:52:51,880 --> 00:52:53,560 BUT WE DON'T FULLY UNDERSTAND 1105 00:52:53,560 --> 00:52:56,760 WHY IT HAS THIS SELECTIVITY. 1106 00:52:56,760 --> 00:52:59,400 WHAT ELSE DOES IT MEDIATE INSIDE 1107 00:52:59,400 --> 00:53:03,400 A CELL? AND FINALLY, OUR 1108 00:53:03,400 --> 00:53:05,400 UNPUBLISHED DATA SHOWING THAT IT 1109 00:53:05,400 --> 00:53:07,160 IS ALSO MODIFIED BY THIS LIPID 1110 00:53:07,160 --> 00:53:10,160 GROUP AND I THINK THIS 1111 00:53:10,160 --> 00:53:16,600 CONNECTION TO ROS ALSO SAYS THAT 1112 00:53:16,600 --> 00:53:17,880 PYROPTOSIS IS NOT JUST ANY FORM 1113 00:53:17,880 --> 00:53:22,840 OF CELL DEATH. IT'S A CELL 1114 00:53:22,840 --> 00:53:25,120 DEATH OF THE LAST RESORT IF YOU 1115 00:53:25,120 --> 00:53:28,080 WILL. THE RESORTS ARE ALREADY 1116 00:53:28,080 --> 00:53:30,560 REALLY STRESSED OUT BY STRESS. 1117 00:53:30,560 --> 00:53:33,400 BY OXIDATIVE STRESS, FOR 1118 00:53:33,400 --> 00:53:37,400 EXAMPLE. AND THEN, YOU KNOW, 1119 00:53:37,400 --> 00:53:39,920 PYROPTOSIS IS ACHIEVED THROUGH 1120 00:53:39,920 --> 00:53:42,960 THIS CONDUIT SO MAYBE PYROPTOSIS 1121 00:53:42,960 --> 00:53:49,200 SHOULD REALLY BE AN OXIDATIVE 1122 00:53:49,200 --> 00:53:51,880 INDUCED CELL DEATH. WITH THAT, 1123 00:53:51,880 --> 00:53:54,560 LET ME THANK THE POST-SECONDARY 1124 00:53:54,560 --> 00:53:56,480 DOC STUDENTS AND I WANT TO 1125 00:53:56,480 --> 00:53:58,240 HIGHLIGHT MY COLLABORATION WITH 1126 00:53:58,240 --> 00:54:00,160 MY COLLEAGUE JULIE LIEBERMAN. 1127 00:54:00,160 --> 00:54:05,120 AND I THANK YOU FOR YOUR 1128 00:54:05,120 --> 00:54:05,400 ATTENTION. 1129 00:54:05,400 --> 00:54:11,440 >> (APPLAUSE). 1130 00:54:11,440 --> 00:54:14,600 >> HELLO, WOW. THANK YOU VERY 1131 00:54:14,600 --> 00:54:16,280 MUCH. LET'S HAVE ANOTHER ROUND 1132 00:54:16,280 --> 00:54:16,640 OF APPLAUSE. 1133 00:54:16,640 --> 00:54:17,360 >> THANK YOU. 1134 00:54:17,360 --> 00:54:21,720 >> THAT'S GREAT. 1135 00:54:21,720 --> 00:54:23,440 >> I DIDN'T GET ANY QUESTIONS ON 1136 00:54:23,440 --> 00:54:25,480 HERE. OH, WAIT, THERE'S ONE 1137 00:54:25,480 --> 00:54:29,440 HERE. SO LET'S -- 1138 00:54:29,440 --> 00:54:29,720 >> OKAY. 1139 00:54:29,720 --> 00:54:33,440 >> JAY, SOME REPORTS HAVE 1140 00:54:33,440 --> 00:54:34,120 INDICATED THAT MITOCHONDRIAL 1141 00:54:34,120 --> 00:54:36,480 DATA RELEASE IS REQUIRED FOR 1142 00:54:36,480 --> 00:54:38,440 NLPR 3 ACTIVATION. DO YOU SEE 1143 00:54:38,440 --> 00:54:43,200 EVIDENCE FOR THIS? 1144 00:54:43,200 --> 00:54:45,280 >> YES, I TALKED TO MICHAEL 1145 00:54:45,280 --> 00:54:48,880 ABOUT THIS PRETTY EXTENSIVELY. 1146 00:54:48,880 --> 00:54:54,720 YEAH. WE TESTED PREVIOUSLY, TO 1147 00:54:54,720 --> 00:54:57,320 OUR INACTIVE TO SEE IF IT BINDS. 1148 00:54:57,320 --> 00:54:59,360 WE DID NOT SEE INTERACTION IN 1149 00:54:59,360 --> 00:55:01,640 OUR HANDS BUT NOW WE HAVE THE 1150 00:55:01,640 --> 00:55:07,760 ACTIVE SATE -- STATE TO SEE IF 1151 00:55:07,760 --> 00:55:08,240 IT BINDS AGAIN. 1152 00:55:08,240 --> 00:55:09,440 I WOULD LIKE TO MAKE A 1153 00:55:09,440 --> 00:55:11,160 CONNECTION BETWEEN THE 1154 00:55:11,160 --> 00:55:13,440 MITOCHONDRIA PATHWAY AND THE 1155 00:55:13,440 --> 00:55:15,360 TGN. I DON'T KNOW HOW TO 1156 00:55:15,360 --> 00:55:16,160 RATIONALIZE THIS WHOLE THING YET 1157 00:55:16,160 --> 00:55:17,720 BUT HOPEFULLY WE'LL GET THERE. 1158 00:55:17,720 --> 00:55:19,400 >> GREAT. OTHER QUESTIONS? 1159 00:55:19,400 --> 00:55:26,040 >>THIS IS PETER. YOU DESCRIBE LUCID 1160 00:55:26,040 --> 00:55:29,080 EXTENSIVELY IN THE VESICLE 1161 00:55:29,080 --> 00:55:32,880 TRAFFICKING AND TGN. BUT THE 1162 00:55:32,880 --> 00:55:36,840 PRIMARY FUNCTION IS ACTIVATE PER 1163 00:55:36,840 --> 00:55:40,000 CASPASE, RIGHT? SO DO YOU SEE 1164 00:55:40,000 --> 00:55:45,640 ANY EVIDENCE? HOW PRO-CASPASE-1 1165 00:55:45,640 --> 00:55:49,240 GETS INTERACT WITH THE 1166 00:55:49,240 --> 00:55:50,080 INFLAMMASOME AND THEN 1167 00:55:50,080 --> 00:55:50,440 SUBSEQUENTLY -- 1168 00:55:50,440 --> 00:55:53,680 >> YEAH. YEAH. YEAH. SO THAT 1169 00:55:53,680 --> 00:55:54,720 ACTUALLY I THINK THAT 1170 00:55:54,720 --> 00:55:57,640 QUESTION -- WELL, IT MAY BE MORE 1171 00:55:57,640 --> 00:56:01,800 COMPLEX BUT IN OUR I THINK 2016 1172 00:56:01,800 --> 00:56:03,600 PAPER OR SOMETHING SO WE LOOKED 1173 00:56:03,600 --> 00:56:06,680 AT HOW IT INTERACT. SO IT HAS A 1174 00:56:06,680 --> 00:56:11,120 DOMAIN ON THE TERMINUS AND 1175 00:56:11,120 --> 00:56:13,400 INTERACT WITH THE C DOMAIN AND 1176 00:56:13,400 --> 00:56:16,440 ONCE THIS INTERACTION OCCURS 1177 00:56:16,440 --> 00:56:17,640 THIS WILL BE BROUGHT TOGETHER 1178 00:56:17,640 --> 00:56:19,720 AND THEN THEY DIE AMOR RISE SO 1179 00:56:19,720 --> 00:56:23,120 THEN THEY CAN THEN 1180 00:56:23,120 --> 00:56:25,400 AUTOPROCESS -- TRANSAUTOPROCESS 1181 00:56:25,400 --> 00:56:27,080 TO BECOME CLEAVED AND ACTIVE. 1182 00:56:27,080 --> 00:56:30,120 SO MANY MY HEAD THIS IS WHAT'S 1183 00:56:30,120 --> 00:56:31,160 HAPPENING. BUT I THINK THE 1184 00:56:31,160 --> 00:56:33,120 STORY PROBABLY IS MUCH MORE 1185 00:56:33,120 --> 00:56:34,120 COMPLEX. WE HAVE SOME 1186 00:56:34,120 --> 00:56:36,880 UNPUBLISHED DATA WHICH IS STILL 1187 00:56:36,880 --> 00:56:38,600 NOT COMPLETELY CLEAR BUT WE 1188 00:56:38,600 --> 00:56:43,400 THINK THAT THE DIRECT 1189 00:56:43,400 --> 00:56:44,840 RECRUITMENT OF PROCYTOKINES, 18 1190 00:56:44,840 --> 00:56:46,680 AND L 1 IS NOT THAT SIMPLE. 1191 00:56:46,680 --> 00:56:49,040 IT'S ACTUALLY, YOU KNOW, IT'S -- 1192 00:56:49,040 --> 00:56:51,640 MORE COMPLEX THAN JUST A SIMPLE 1193 00:56:51,640 --> 00:56:52,240 RECRUITMENT. WE THINK THERE'S 1194 00:56:52,240 --> 00:56:54,440 MORE STEPS THAT'S INVOLVED IN 1195 00:56:54,440 --> 00:56:56,640 THERE BUT WE DON'T KNOW ABOUT 1196 00:56:56,640 --> 00:56:58,080 THE SUBSTRATE RECOGNITION YET 1197 00:56:58,080 --> 00:57:00,800 BUT I THINK WE'RE PRETTY CLEAR 1198 00:57:00,800 --> 00:57:02,600 ABOUT THE CASPASE ACTIVATION BUT 1199 00:57:02,600 --> 00:57:05,080 WE DON'T KNOW ABOUT SUBSTRATE 1200 00:57:05,080 --> 00:57:06,840 RECRUITMENT. AND AS YOU KNOW, 1201 00:57:06,840 --> 00:57:11,760 FUNCTIONAL GROUP AND ALSO I 1202 00:57:11,760 --> 00:57:13,040 THINK THE GROUP HAVE SHOWN 1203 00:57:13,040 --> 00:57:16,760 THAT -- HAS SHOWN THAT GASDERMIN 1204 00:57:16,760 --> 00:57:21,520 D NEEDS THIS SITE TO BE 1205 00:57:21,520 --> 00:57:24,000 RECRUITED TO CASPASE-1. SO IT'S 1206 00:57:24,000 --> 00:57:27,640 NOT JUST AN ACTIVE SITE 1207 00:57:27,640 --> 00:57:29,040 INTERACTION BUT ALSO AN ACYL 1208 00:57:29,040 --> 00:57:31,400 SITE INTERACTION. THIS IS MORE 1209 00:57:31,400 --> 00:57:34,560 IMPORTANT THAN THE MOTIF 1210 00:57:34,560 --> 00:57:37,400 RECOGNITION. SO I -- I THINK 1211 00:57:37,400 --> 00:57:41,000 OUR DATA MAY BEGIN TO SHOW THAT 1212 00:57:41,000 --> 00:57:45,040 THE -- SOME KIND OF AN XO SITE 1213 00:57:45,040 --> 00:57:47,040 INTERACTION IS ALSO FOR 1214 00:57:47,040 --> 00:57:47,760 PROCYTOKINE PROCESSING. 1215 00:57:47,760 --> 00:57:50,960 >> MY UNDERSTANDING IS THAT 1216 00:57:50,960 --> 00:57:52,560 ACTIVATION, CASPASE DOES REQUIRE 1217 00:57:52,560 --> 00:57:54,760 ASSEMBLY OF INFLAM SOME SO IT 1218 00:57:54,760 --> 00:57:59,760 POSSIBLE TO PUT A CATALYTICALLY 1219 00:57:59,760 --> 00:58:00,480 DEAD CASPASE CONSTRUCT INTO 1220 00:58:00,480 --> 00:58:03,280 YOUR -- ANY. 1221 00:58:03,280 --> 00:58:06,080 >> YES, ABSOLUTELY, YEAH. M-HM. 1222 00:58:06,080 --> 00:58:08,000 WE THINK THE DIE AMORIZATION 1223 00:58:08,000 --> 00:58:09,080 WILL STILL OCCUR WITHIN THE 1224 00:58:09,080 --> 00:58:09,800 FILAMENT, YEAH. 1225 00:58:09,800 --> 00:58:12,600 >> YES. 1226 00:58:12,600 --> 00:58:15,200 >> THANK YOU SO MUCH FOR 1227 00:58:15,200 --> 00:58:21,040 WONDERFUL TALK, DR. WU. I AM 1228 00:58:21,040 --> 00:58:24,640 ACTUALLY FASCINATED ABOUT THE 1229 00:58:24,640 --> 00:58:27,880 PAL MY TOLATION AND YOUR DATA 1230 00:58:27,880 --> 00:58:28,840 BEAUTIFULLY EXPLAINS WHAT 1231 00:58:28,840 --> 00:58:30,920 HAPPENS WHEN IT IS ACTIVATED. I 1232 00:58:30,920 --> 00:58:33,080 WOULD LIKE TO ASK IN CASE OTHER 1233 00:58:33,080 --> 00:58:35,520 INFLAMMASOMES ARE ACTIVATED. DO 1234 00:58:35,520 --> 00:58:39,880 YOU THINK THE SAME THING OCCURS 1235 00:58:39,880 --> 00:58:41,400 TO GASDERMIN M.D. BECAUSE -- 1236 00:58:41,400 --> 00:58:43,320 >> WE HAVEN'T TESTED BUT I THINK 1237 00:58:43,320 --> 00:58:45,080 SO. I THINK BECAUSE THIS IS A 1238 00:58:45,080 --> 00:58:46,680 PHYSICAL PRINCIPLE. SO I DON'T 1239 00:58:46,680 --> 00:58:48,520 THINK IT'LL BE DIFFERENT. 1240 00:58:48,520 --> 00:58:50,760 UNLESS THERE'S OTHER MECHANISMS 1241 00:58:50,760 --> 00:58:57,040 TO COMPENSATE FOR THAT. I 1242 00:58:57,040 --> 00:59:01,360 IMAGINE OUR INFLAMMASOME 1243 00:59:01,360 --> 00:59:03,720 ACTIVATION RAISE ROS. I DON'T 1244 00:59:03,720 --> 00:59:09,040 KNOW HOW AND THAT WILL CAUSE 1245 00:59:09,040 --> 00:59:12,160 GASDERMIN M.D. AT LEAST THAT'S 1246 00:59:12,160 --> 00:59:13,480 WHAT I IMAGINE. 1247 00:59:13,480 --> 00:59:16,120 BUT WE HAVEN'T TESTED THAT. 1248 00:59:16,120 --> 00:59:17,280 >> OKAY. ANOTHER QUESTION, 1249 00:59:17,280 --> 00:59:22,720 IS -- DO YOU THINK THAT IN OTHER 1250 00:59:22,720 --> 00:59:24,880 GASDERMINS, IS THE CYSTEINE 1251 00:59:24,880 --> 00:59:25,960 RESIDUE CONSERVED. 1252 00:59:25,960 --> 00:59:28,880 >> NO, IT IS NOT CONSERVED BUT 1253 00:59:28,880 --> 00:59:32,520 OTHER GASDERMINS THAT WE'RE ABLE 1254 00:59:32,520 --> 00:59:34,800 TO TEST SO FAR ARE 1255 00:59:34,800 --> 00:59:35,720 PALMITOYLATED. THERE'S ONLY ONE 1256 00:59:35,720 --> 00:59:37,040 WE COULDN'T TEST BECAUSE IT 1257 00:59:37,040 --> 00:59:39,040 DIDN'T EXPRESS WHICH IS 1258 00:59:39,040 --> 00:59:41,000 GASDERMIN C BUT EVERYTHING ELSE 1259 00:59:41,000 --> 00:59:44,920 WE TESTED OR PALMITOYLATED. AS 1260 00:59:44,920 --> 00:59:47,120 A STRUCTURALIST I LIKE SIMPLE 1261 00:59:47,120 --> 00:59:49,440 THINGS. SO I THINK THEY -- 1262 00:59:49,440 --> 00:59:52,160 THERE'S GOT TO BE SOME 1263 00:59:52,160 --> 00:59:52,720 COMMONALITY AMONG ALL THE 1264 00:59:52,720 --> 00:59:53,880 DIFFERENT GASDERMINS. 1265 00:59:53,880 --> 00:59:57,040 >> OKAY. THANK YOU. 1266 00:59:57,040 --> 00:59:57,640 >> YES. 1267 00:59:57,640 --> 01:00:02,200 >> VERY NICE TALK WITH THE NLRP 1268 01:00:02,200 --> 01:00:05,040 3 SUMMARY SLIDE YOU SHOW THAT 1269 01:00:05,040 --> 01:00:08,080 HAD THE INHIBITED BUT CAGED FORM 1270 01:00:08,080 --> 01:00:09,840 IS WHAT GETS TRAFFICKED. HOW 1271 01:00:09,840 --> 01:00:11,760 DOES THAT HAPPEN? WHY DOES IT 1272 01:00:11,760 --> 01:00:13,440 HAVE TO BE IN THAT CAGED 1273 01:00:13,440 --> 01:00:16,040 INHIBITED FORM? HOW IS THE 1274 01:00:16,040 --> 01:00:17,080 SELECTIVITY ENFORCED? 1275 01:00:17,080 --> 01:00:19,760 >> WE ARE TRYING TO FIGURE THAT 1276 01:00:19,760 --> 01:00:21,840 OUT. WE DON'T KNOW. SO I THINK 1277 01:00:21,840 --> 01:00:25,440 THE CAGE IS REQUIRED FOR THE 1278 01:00:25,440 --> 01:00:27,680 GOLGI BINDING BECAUSE YOU HAVE 1279 01:00:27,680 --> 01:00:30,240 MULTIVA LENIENCY. IF YOU JUST 1280 01:00:30,240 --> 01:00:32,560 HAVE ONE, YOU KNOW, POSITIVELY 1281 01:00:32,560 --> 01:00:33,360 CHARGED PATCH IT'S PROBABLY NOT 1282 01:00:33,360 --> 01:00:36,520 ENOUGH YOU NEED THE WHOLE THING. 1283 01:00:36,520 --> 01:00:39,400 SO BUT WHY DOES THE CAGE INDUCE 1284 01:00:39,400 --> 01:00:40,760 GOLGI DISPERSION? THAT WE DON'T 1285 01:00:40,760 --> 01:00:42,400 KNOW. AND THAT'S A QUESTION WE 1286 01:00:42,400 --> 01:00:47,080 REALLY WANT TO ADDRESS, YEAH. 1287 01:00:47,080 --> 01:00:50,240 >> OKAY. I'M POSTDOC IN MAX 1288 01:00:50,240 --> 01:00:54,400 LAB. SO I'M WONDERING IF 1289 01:00:54,400 --> 01:00:58,640 GASDERMIN D IS ALMOST SELECTIVE 1290 01:00:58,640 --> 01:00:59,520 FOR MEMORY. 1291 01:00:59,520 --> 01:01:00,920 >> I DIDN'T MENTION THAT BUT 1292 01:01:00,920 --> 01:01:03,920 CERTAINLY MITOCHONDRIA. 1293 01:01:03,920 --> 01:01:06,800 >> TWO LAYER MEMORY. 1294 01:01:06,800 --> 01:01:08,240 >> YEAH. SO THERE'S -- MAYBE 1295 01:01:08,240 --> 01:01:12,800 THOSE ARE NOT PUBLISHED BUT I 1296 01:01:12,800 --> 01:01:15,360 THINK MITOCHONDRIA DAMAGE BY 1297 01:01:15,360 --> 01:01:16,920 GASDERMIN HAS BEEN IMPLICATED. 1298 01:01:16,920 --> 01:01:18,920 MAYBE NOT DIRECTLY SHOWN BUT 1299 01:01:18,920 --> 01:01:21,840 IMPLICATED. YEAH. SO THE IDEA 1300 01:01:21,840 --> 01:01:25,440 IS THAT THE LIKING THAT'S IN THE 1301 01:01:25,440 --> 01:01:27,400 MEMBRANE OF THE MITOCHONDRIA CAN 1302 01:01:27,400 --> 01:01:29,680 BE EXTERNALIZED TO THE OUTSIDE. 1303 01:01:29,680 --> 01:01:31,840 IN FACT, THE SCRAMBLING THAT'S 1304 01:01:31,840 --> 01:01:35,760 IMPORTANT FOR THIS PROCESS IS 1305 01:01:35,760 --> 01:01:38,080 IMPORTANT FOR GASDERMIN D 1306 01:01:38,080 --> 01:01:39,360 LOCALIZATION TO THE 1307 01:01:39,360 --> 01:01:39,720 MITOCHONDRIA. 1308 01:01:39,720 --> 01:01:42,440 >> ANOTHER QUESTION IS AS YOU 1309 01:01:42,440 --> 01:01:44,640 SAID, THE GASDERMIN D BE CLEAVED 1310 01:01:44,640 --> 01:01:47,600 INTO AN N TERMINAL. WHAT IS THE 1311 01:01:47,600 --> 01:01:47,840 FUNCTION. 1312 01:01:47,840 --> 01:01:49,400 >> I HAVE NO IDEA WHAT THE C 1313 01:01:49,400 --> 01:01:50,080 TERMINAL DOES. I HAVE TO IDEA. 1314 01:01:50,080 --> 01:01:53,040 YEAH. YEAH. 1315 01:01:53,040 --> 01:01:57,080 >> DO THEY FORM, LIKE, A DIMER 1316 01:01:57,080 --> 01:01:59,520 OR ONLY THE -- THE C TERMINAL 1317 01:01:59,520 --> 01:02:01,080 RELEASE AND THE FUNCTION -- 1318 01:02:01,080 --> 01:02:05,080 >> YEAH, WE DON'T -- SO THE ONLY 1319 01:02:05,080 --> 01:02:07,640 FUNCTION I KNOW ABOUT FOR THE C 1320 01:02:07,640 --> 01:02:10,120 TERMINUS IS TO SUPPRESS THE N 1321 01:02:10,120 --> 01:02:11,200 TERMINUS FOR SURE IN THE PROTEIN 1322 01:02:11,200 --> 01:02:13,440 IT DOES THAT. ONCE IT'S 1323 01:02:13,440 --> 01:02:15,400 RELEASED WHAT DOES IT DO? I 1324 01:02:15,400 --> 01:02:17,480 HAVE NO CLUE. I HAVE NO IDEA 1325 01:02:17,480 --> 01:02:21,040 WHAT IT DOES AFTERWARDS: YEAH, 1326 01:02:21,040 --> 01:02:23,720 MAYBE IT'S NOT FULLY -- OH, SO I 1327 01:02:23,720 --> 01:02:28,440 WANT TO MENTION ONE THING. SO 1328 01:02:28,440 --> 01:02:29,320 GASDERMIN D I DIDN'T TALK ABOUT 1329 01:02:29,320 --> 01:02:31,360 THAT PART OF OUR WORK BUT 1330 01:02:31,360 --> 01:02:33,840 GASDERMIN D IS EXPRESSED WIDELY 1331 01:02:33,840 --> 01:02:36,840 IN CELLS. NOT JUST IN 1332 01:02:36,840 --> 01:02:38,720 MACROPHAGES SO THAT WAS REALLY 1333 01:02:38,720 --> 01:02:41,200 STRANGE. IF IT'S DOWNSTREAM OF 1334 01:02:41,200 --> 01:02:42,280 INFLAMMASOME WHY IS IT EXPRESSED 1335 01:02:42,280 --> 01:02:44,760 IN OTHER CELLS? FOR EXAMPLE, IN 1336 01:02:44,760 --> 01:02:48,520 A LOT OF CANCER CELLS IT'S 1337 01:02:48,520 --> 01:02:51,400 EXPRESSED. SO I DON'T KNOW, BUT 1338 01:02:51,400 --> 01:02:52,600 I IMAGINE GASDERMIN D HAS OTHER 1339 01:02:52,600 --> 01:02:55,400 FUNCTIONS. MAYBE THAT WILL HAVE 1340 01:02:55,400 --> 01:02:57,000 TO DO WITH THE C TERMINAL 1341 01:02:57,000 --> 01:02:58,200 DOMAIN. IT'S ANYONE'S GUESS I 1342 01:02:58,200 --> 01:03:01,760 THINK AT THIS POINT. YEAH. 1343 01:03:01,760 --> 01:03:03,400 >> SO THE CLEAVE, THE GASDERMIN 1344 01:03:03,400 --> 01:03:04,680 D IS ONLY ONE -- 1345 01:03:04,680 --> 01:03:09,080 >> NO, THERE'S ALSO OTHER ONES. 1346 01:03:09,080 --> 01:03:12,040 >> LIKE RUN DOWN SOME. 1347 01:03:12,040 --> 01:03:12,920 >> DEPENDS ON WHICH ONE. B I 1348 01:03:12,920 --> 01:03:18,160 THINK. NO, THAT'S CLEAVING -- 1349 01:03:18,160 --> 01:03:19,760 THAT'S ACTUALLY CLEAVING E. 1350 01:03:19,760 --> 01:03:21,400 IT'S BETTER I GO THROUGH MY 1351 01:03:21,400 --> 01:03:23,800 TALK. I ACTUALLY DON'T REMEMBER 1352 01:03:23,800 --> 01:03:29,040 THE C. OH -- I DON'T KNOW HOW 1353 01:03:29,040 --> 01:03:39,600 TO GO BACK, OKAY. SO IT'S HERE. 1354 01:03:40,480 --> 01:03:44,120 CASPASE A ALSO CLEAVES D. AND 1355 01:03:44,120 --> 01:03:47,120 THE KNEW TRUE FILL ELASTASE 1356 01:03:47,120 --> 01:03:48,840 CLOOEAGE OF D IS ACTUALLY 1357 01:03:48,840 --> 01:03:52,080 IMPORTANT FOR ME TO SIS. THIS 1358 01:03:52,080 --> 01:03:55,360 NEUTROPHIL EXCLUSION OF NUCLEAR 1359 01:03:55,360 --> 01:03:59,200 DNA AND CASPASE A THROUGH THE 1360 01:03:59,200 --> 01:04:03,760 APOPTOTIC PATHWAY CAN ALSO 1361 01:04:03,760 --> 01:04:04,200 CLEAVE GASDERMIN D. 1362 01:04:04,200 --> 01:04:04,600 >> THANK YOU. 1363 01:04:04,600 --> 01:04:05,040 >> YEAH. 1364 01:04:05,040 --> 01:04:06,880 >> (SPEAKER FAR FROM MIC). 1365 01:04:06,880 --> 01:04:08,840 >> YEAH. IT'S ABOUT -- I 1366 01:04:08,840 --> 01:04:11,440 MEASURED THIS BEFORE AND NOW I 1367 01:04:11,440 --> 01:04:12,680 DON'T KNOW IF I REMEMBER WHEN I 1368 01:04:12,680 --> 01:04:15,480 WAS WRITING THE FIRST PAPER I 1369 01:04:15,480 --> 01:04:17,480 MEASURED IT. IT'S ABOUT A 1370 01:04:17,480 --> 01:04:19,440 SLIGHTLY COMPRESSED DOUBLE LA 1371 01:04:19,440 --> 01:04:23,200 LAYER. IT'S LIKE SLIGHTLY 1372 01:04:23,200 --> 01:04:24,800 COMPRESSED I WOULD SAY. SO IT'S 1373 01:04:24,800 --> 01:04:29,040 POSSIBLE IT CAUSES SOME MEMBRANE 1374 01:04:29,040 --> 01:04:29,440 CURVATURE AS WELL. 1375 01:04:29,440 --> 01:04:35,000 >> (SPEAKER FAR FROM MIC). 1376 01:04:35,000 --> 01:04:36,600 >> IT IS GOING THROUGH THE 1377 01:04:36,600 --> 01:04:39,240 MEMBRANE ALREADY. IF YOU CHANGE 1378 01:04:39,240 --> 01:04:40,800 THE HYDROPHOBIC BITS. YEAH. 1379 01:04:40,800 --> 01:04:42,720 I'VE NOT DONE THAT. YOU WONDER 1380 01:04:42,720 --> 01:04:44,080 WHETHER THAT WILL CHANGE THE 1381 01:04:44,080 --> 01:04:46,520 SELECTIVITY. YEAH. IT MAY. 1382 01:04:46,520 --> 01:04:46,720 YEAH. 1383 01:04:46,720 --> 01:04:47,520 >> ONE MORE QUESTION? 1384 01:04:47,520 --> 01:04:48,640 >> ONE MORE FINAL QUESTION. 1385 01:04:48,640 --> 01:04:51,720 >> THANK YOU FOR THE TALK. I 1386 01:04:51,720 --> 01:04:56,560 HAVE ONE QUESTION. WHY THE 1387 01:04:56,560 --> 01:04:59,360 SAMPLE TEST? I WANT TO KNOW 1388 01:04:59,360 --> 01:04:59,560 THE -- 1389 01:04:59,560 --> 01:05:00,720 >> YEAH. YEAH. 1390 01:05:00,720 --> 01:05:05,600 >> THE MOLECULAR BASIS OF THE 1391 01:05:05,600 --> 01:05:06,400 FUNCTION. 1392 01:05:06,400 --> 01:05:08,920 >> YEAH. YEAH. SO WE FOUND THE 1393 01:05:08,920 --> 01:05:11,400 REASON THAT WE INVESTIGATED WAS 1394 01:05:11,400 --> 01:05:12,040 BECAUSE WE SEE THESE SINGLE 1395 01:05:12,040 --> 01:05:14,440 SPECS IN A CELL. WE WERE, LIKE, 1396 01:05:14,440 --> 01:05:17,040 WHY? WHY IS IT ONLY ONE? WHY 1397 01:05:17,040 --> 01:05:19,440 IS EVERYTHING COALESCE TO ONE? 1398 01:05:19,440 --> 01:05:22,600 AND IT'S PARA NUCLEAR SO WE WERE 1399 01:05:22,600 --> 01:05:26,760 JUST, LIKE, IT LOOKS LIKE MTOC 1400 01:05:26,760 --> 01:05:30,000 SO WE TESTED BY 1401 01:05:30,000 --> 01:05:31,520 IMMUNOFLUORESCENCE AND INDEED 1402 01:05:31,520 --> 01:05:36,720 THEY ARE ONLY MTOC AND THEN WE 1403 01:05:36,720 --> 01:05:41,400 TESTED MICROTUBULE SENSITIVITY, 1404 01:05:41,400 --> 01:05:45,000 ARE THESE THINGS SENSITIVE TO 1405 01:05:45,000 --> 01:05:46,200 MICROTUBULE STRUCTURES AND THIS 1406 01:05:46,200 --> 01:05:51,280 IS ALSO BEFORE US. EVEN JURG 1407 01:05:51,280 --> 01:05:52,920 TESTED THESE AND THEY ALL 1408 01:05:52,920 --> 01:05:55,440 DISRUPT THE ACTIVATION SO WE 1409 01:05:55,440 --> 01:05:58,240 RETESTED THOSE AND ALSO MAPPED 1410 01:05:58,240 --> 01:06:00,800 THE INHIBITOR ALSO INHIBITED IN 1411 01:06:00,800 --> 01:06:04,080 RP 3 AND THE ADAPTER ADAPTER 1412 01:06:04,080 --> 01:06:10,560 WHICH IS HDAK 6 WHICH BINDS ALSO 1413 01:06:10,560 --> 01:06:18,760 INHIBIT THE MICROTUBULE 1414 01:06:18,760 --> 01:06:19,560 TRANSPORT. WHAT'S THE REASON 1415 01:06:19,560 --> 01:06:21,920 FOR THAT? GOOD QUESTION. MY 1416 01:06:21,920 --> 01:06:23,640 RATIONALE FOR THAT IS THIS 1417 01:06:23,640 --> 01:06:29,040 PATHWAY IS ALSO SIMILAR TO THE 1418 01:06:29,040 --> 01:06:32,720 SO-CALLI 1419 01:06:32,720 --> 01:06:33,280 SO-CALLING A ING A RAH SOME 1420 01:06:33,280 --> 01:06:33,520 PATHWAY. 1421 01:06:33,520 --> 01:06:36,200 THEY ARE TOO BIG I THINK TO BE 1422 01:06:36,200 --> 01:06:37,800 DEGRADED BY THE PROTEOSOME SO 1423 01:06:37,800 --> 01:06:41,720 THEY GET TRANSPORTED TO THE MTOC 1424 01:06:41,720 --> 01:06:43,080 AND AT THE MTOC THERE'S USUALLY 1425 01:06:43,080 --> 01:06:44,800 A LOT OF DONOR MEMBRANES. YOU 1426 01:06:44,800 --> 01:06:46,120 HAVE MITOCHONDRIA RIGHT THERE. 1427 01:06:46,120 --> 01:06:49,000 THERE'S LOTS OF THINGS THERE. 1428 01:06:49,000 --> 01:06:51,080 SO YOU FORM OTHERS. SO THAT'S 1429 01:06:51,080 --> 01:06:53,040 HOW YOU DEGRADE THESE PROTEINS 1430 01:06:53,040 --> 01:06:55,520 SO IN OUR CASE, WE ALSO SAW THE 1431 01:06:55,520 --> 01:06:58,480 SPEC. THIS WAS THE OLD WORK IN 1432 01:06:58,480 --> 01:07:01,480 OUR SPEC THEY'RE ENCLOSED 1433 01:07:01,480 --> 01:07:07,400 OFTENTIMES BY DOUBLE MEMBRANES. 1434 01:07:07,400 --> 01:07:11,200 SO WE THINK IT GOES THERE TO 1435 01:07:11,200 --> 01:07:15,000 REGULATE AND IF WE TREAT WITH 1436 01:07:15,000 --> 01:07:19,320 3MA THAT INHIBIT THE PUMP THAT 1437 01:07:19,320 --> 01:07:21,320 GOES FROM HERE TO HERE ARE HUGE. 1438 01:07:21,320 --> 01:07:27,120 THEY HAVE THIS GIGANTIC AND IT'S 1439 01:07:27,120 --> 01:07:29,440 AT THE LEVEL OF INFLAMMASOME 1440 01:07:29,440 --> 01:07:31,360 ACTIVATION OF THE CELL. SO WE 1441 01:07:31,360 --> 01:07:33,160 THINK WE GO THERE TO ACTIVATE 1442 01:07:33,160 --> 01:07:35,000 AND GO THERE TO BE REGULATED. 1443 01:07:35,000 --> 01:07:37,440 >> YES. THANK YOU. I THINK YOU 1444 01:07:37,440 --> 01:07:41,040 ARE -- THERE SHOULD BE A 1445 01:07:41,040 --> 01:07:44,960 MECHANISM RECRUITING THE RP 3 OR 1446 01:07:44,960 --> 01:07:47,640 THE MECHANISM TO THE MTOC. I 1447 01:07:47,640 --> 01:07:48,720 THINK THERE SHOULD BE A 1448 01:07:48,720 --> 01:07:49,440 MECHANISM THERE. 1449 01:07:49,440 --> 01:07:53,040 >> WE THINK IT'S ON THE VESICLE. 1450 01:07:53,040 --> 01:07:55,360 THAT'S WHAT WE THINK. THE 1451 01:07:55,360 --> 01:07:56,600 DISPERSION. WE THINK IT'S THE 1452 01:07:56,600 --> 01:07:58,000 VESICLES THAT GETS TRANSPORTED 1453 01:07:58,000 --> 01:08:00,040 BUT WE'RE NOT 100% SURE. THE 1454 01:08:00,040 --> 01:08:02,840 REASON WE SAID IT'S THE VESICLE 1455 01:08:02,840 --> 01:08:06,000 THAT GETS TRANSPORTED IS BECAUSE 1456 01:08:06,000 --> 01:08:07,680 TGM MARKERS BECOME ON THE MTOC 1457 01:08:07,680 --> 01:08:10,240 WHEN YOU HAVE INFLAM SOMES SO WE 1458 01:08:10,240 --> 01:08:12,520 FIGURED THAT THAT MUST BE THE 1459 01:08:12,520 --> 01:08:17,840 VESICLE THAT GETS TRANSPORTED. 1460 01:08:17,840 --> 01:08:18,760 >> THANK YOU VERY MUCH. THAT 1461 01:08:18,760 --> 01:08:20,520 WAS REALLY FANTASTIC. 1462 01:08:20,520 --> 00:00:00,000 >> THANK YOU.