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è¿‘æ—¥ï¼?/span>北京大å¦åŒ–å¦ä¸Žåˆ†å工程å¦é™¢ã€?/span>北大-清åŽç”Ÿå‘½ç§‘å¦è”åˆä¸å¿ƒé™ˆé¹æ•™æŽˆè¯¾é¢˜ç»?/span>以â€?/span>Enzyme-Mediated Intercellular Proximity Labeling for Detecting Cell–Cell Interactionsâ€?/span>为题åœ?/span>J. Am. Chem. Soc.æ‚志报é“了一ç§èƒ½å¤Ÿåœ¨æ´»ä½“å’?/span>åŽŸä½æƒ…况ä¸?/span>æ•获细胞之间相互作用的技æœ?/span>ï¼?/span>美国化å¦ä¼?/span>C&E NEWSäº?/span>2æœ?/span>12日以â€?/span>New labeling method can reveal cell-cell interactionsâ€?/span>为题进行了专题报é“ã€?/span>
ä¸åŒç»†èƒžä¹‹é—´çš„相互作用和信æ¯ä¼ 递在包括å…ç–«å“应ã€?/span>器官å‘育ã€ç¥žç»ä¼ å¯?/span>在内的众多生命活动ä¸éƒ½æ‰®æ¼?/span>ç€å…³é”®è§’色。目å‰ï¼Œç ”究细胞互作的方æ³?/span>å¤?/span>éƒ?/span>需è¦?/span>已知å‚与çš?/span>ç»†èƒžç±»åž‹ï¼Œéš¾ä»¥åœ¨å¤æ‚的活体环境下å‘çŽ°æˆ–ç ”ç©¶æœªçŸ¥çš„ç»†èƒžé—?/span>相互作用ã€?/span>为了解决这一难题,陈é¹è¯¾é¢˜ç»„借助“定å‘è¿›åŒ–â€æŠ€æœ¯å’Œâ€?/span>é‚»è¿‘æ ‡è®°â€ç–略,实现äº?/span>细胞之间动æ€ç›¸äº’ä½œç”¨çš„åŽŸä½æ•获。这一被命å为EXCELLï¼?/span>Enzyme-mediated proximal cell labelingï¼?/span>的技术,通过å°?/span>作è€?/span>定å‘进化得到的分选酶ï¼?/span>mgSrtAï¼?/span>展示åœ?/span>å¾…ç ”ç©¶çš„ç»†èƒžè¡¨é¢ï¼?/span>能够对与å…?/span>相互作用çš?/span>ç»†èƒžè¿›è¡ŒåŽŸä½æ•获与鉴å®?/span>ï¼Œä¸ºç ”ç©¶ç»†èƒž-细胞ç›?/span>互作用æä¾›äº†æœ‰åŠ›çš„å·¥å…?/span>ã€?/span>
å›?/span> 1 EXCELL方法用于æ•获和鉴å®?/span>细胞间相互作ç”?/span>
æºè‡ªé‡‘黄色葡è„çƒè?/span>çš?/span>分选酶Sortase A ï¼?/span>SrtAï¼?/span>能够催化分选肽ï¼?/span>LPETGï¼?/span>和寡èšç”˜æ°¨é…¸çš?/span>共价连接。近期有文献报é“å°?/span>SrtA用于监测å°é¼ 体内特定å—体-é…体介导的å…ç–?/span>细胞互作过程1。但是该方法需è¦å°†SrtAä¸?/span>寡èšç”˜æ°¨é…¸åˆ†åˆ«èžåˆåœ¨ç›¸äº’作用çš?/span>é…体和å—ä½?/span>ç»†èƒžä¸Šï¼Œå› æ¤éœ€è¦é¢„çŸ?/span>相互作用çš?/span>细胞类型,并对两类细èƒ?/span>åŒæ—¶è¿›è¡ŒåŸºå› 改é€?/span>ï¼Œå› æ¤æ— æ³•æ•æ‰æœªçŸ¥çš„细胞间相互作ç”?/span>ã€?/span>为了çªç ´è¿™ä¸€ç“¶é¢ˆï¼?/span>陈é¹è¯¾é¢˜ç»?/span>设想获得一ç§é’ˆå¯¹æœªçŸ¥ç»†èƒžçš„æ ‡è®°æ–¹æ³•。由äº?/span>细胞表é¢è›‹ç™½ä¸æœ‰å¤§çº¦5ï¼…å«æœ?/span>N端å•一甘氨é…?/span>ï¼Œå—æ?/span>å¯å‘ï¼?/span>他们希望获得一ç§å¯¹å•个甘氨酸的Nç«¯æ°¨åŸºè¿›è¡Œæ ‡è®°çš„åˆ†é€‰é…¶ï¼?/span>从而能够对任æ„ç»†èƒžåŠ ä»¥æ ‡è®°ã€‚ä¸ºæ?/span>他们建立äº?/span>å¯?/span>SrtA进行定å‘进化的高通é‡è§å…‰ç›é€?/span>å¹³å°ï¼Œå¹¶æˆåŠŸèŽ·å¾—äº?/span>对å•ä¸€ç”˜æ°¨é…¸è¿›è¡Œæ ‡è®°çš„é«˜æ´»æ€?/span>SrtAçªå˜ä½?/span>-mgSrtAï¼?/span>能够实现å¯?/span>ä»»æ„ç»†èƒžç±»åž‹çš„æœ‰æ•ˆæ ‡è®°ã€?/span>
作者分别借助亲和ä½?/span>-抗原ï¼?/span>ZHERä¸?/span>Her2)和é…体-å—体ï¼?/span>CTLA4ä¸?/span>B7, CD40Lä¸?/span>CD40)的相互作用æ?/span>验è¯è¯¥æ–¹æ³?/span>çš„å¯é æ€?/span>ã€?/span>å®žéªŒè¯æ˜Žå±•示在细胞表é¢çš„mgSrtAå?/span>借助邻近效应ï¼?/span>将嫿œ?/span> biotinæ ‡ç¾çš„分选肽biotin-LPETGå…±ä»·æ ‡è®°åœ?/span>与其相互作用çš?/span>细胞表é¢ã€‚通过æµå¼ç»†èƒžæœ?/span>检æµ?/span>ï¼?/span>能够å‘现biotinæ ‡è®°ä¿¡å·ä»…在相互作用细胞上有显著性增强,由æ¤å?/span>æˆåŠŸåŒºåˆ†å’Œæ•获å‘生相互作用的细胞ã€?/span>
å›?/span> 2在两类细胞相互作用模型上应用EXCELL,å¯å®žçŽ°å¯¹äº’ä½?/span>ç»†èƒžçš„æœ‰æ•ˆæ ‡è®?/span>
该方法首次将SrtAæ”¹é€ æˆé«˜æ´»æ€§ä¸”具有æ›?/span>宽泛底物è°?/span>çš?/span>â€?/span>é‚»è¿‘æ ‡è®°â€é…¶ï¼Œå¹¶éªŒè¯å…¶å¯¹ç»†èƒžç›¸äº’ä½œç”¨çš„æ ‡è®°å’Œè¿½è¸ªã€?/span>æ–‡ä¸æŒ‡å‡ºï¼?/span>EXCELLæ–¹æ³•æ— éœ€å°†å¯¡èšç”˜æ°¨é…¸äº‹å…ˆå¼•入到对象细胞表é¢ï¼Œå› æ¤é€‚ç”¨äºŽç ”ç©?/span>夿‚体系ä¸?/span>未知细胞类型çš?/span>相互作用ã€?/span>åŒæ—¶ï¼Œé‰´äº?/span>SrtA底物分选肽ä»?/span>å?/span>五个氨基酸,并已æˆåŠŸåº”ç”¨äºŽå°é¼ 体å†?/span>ã€‚è¯¥æ ‡è®°æ–¹æ³•ä¸ä»…易于监测,在未æ¥è¿˜å¯ä»¥ä¸Žå¤šç§ä¸‹æ¸¸åˆ†æžå¹³å°å…¼å®¹ï¼Œå®Œæˆå¯¹ç›¸äº’作用细胞的æ•获ã€å¯Œé›†å’Œæµ‹åºï¼?/span>从而进一æ¥å¾—出细胞间相互作用在时间ã€ç©ºé—´ã€?/span>类型ç‰å¤šæ–¹é¢çš„ä¿¡æ?/span>ã€?/span>
è¿‘å¹´æ¥ï¼Œä¸€ç³»åˆ—è›‹ç™½è´¨é‚»è¿‘æ ‡è®°æŠ€æœ¯è¢«å¼€å‘并应用äº?/span>蛋白è´?/span>组å¦ç ”ç©¶, æ¤æ¬¡å¼€å‘çš„EXCELL技æœ?/span>实现äº?/span>ç»†èƒžäº’ä½œçš„åŽŸä½æ ‡è®°ä¸Žæ?/span>èŽ?/span>ï¼Œå°†é‚»è¿‘æ ‡è®°ç–略从“胞内â€?/span>拓展è‡?/span>“胞外â€ç©ºé—?/span>ï¼?/span>将在å…疫应ç”ã€ç¥žç»ä¼ 导ç‰é¢†åŸŸå…·æœ‰å¹¿é˜”çš?/span>åº”ç”¨å‰æ™¯ã€?/span>é™ˆé¹æ•™æŽˆä¸ºè¯¥è®?/span>文的通讯作è€?/span>ï¼?/span>课题组åšå£«æ¯•业生葛韵ã€é™ˆé¾™ä¸ºå…±åŒç¬¬ä¸€ä½œè€…。该工作得到了国家自ç„?/span>ç§‘å¦åŸºé‡‘å§”ã€ç§‘技部和北京大å¦åˆæˆä¸ŽåŠŸèƒ½ç”Ÿç‰©åˆ†åä¸å¿?/span>çš?/span>资助ã€?/span>
原文链接�/span>
//pubs.acs.org.ccindex.cn/doi/10.1021/jacs.8b10286
C&E NEWS链接�/span>
//cen.acs.org/biological-chemistry/biochemistry/New-labeling-method-reveal-cell/97/web/2019/02