ÉúÎﻯѧÂÛÎÄ

Äúµ±Ç°µÄλÖãºÑ§ÊõÌà > ÉúÎïѧÂÛÎÄ > ÉúÎﻯѧÂÛÎÄ >

ÐÂÉú³Éµ°°×ºÍÐÞÊε°°×µÄ¼ì²â¼¼Êõ×ÛÊö

À´Ô´£º·ÖÎö»¯Ñ§ ×÷ÕߣºÕ¼·½Áá;¸ß˼Óî;лԪ¶°
·¢²¼ÓÚ£º2020-05-06 ¹²10202×Ö

¡¡¡¡Õª    Òª£º¡¡Ò»¼ÛÍ­´ß»¯µÄµþµªÈ²»ùµÄÌØÒìÐÔ·´Ó¦Í¨³£±»³ÆÎª“µã»÷»¯Ñ§·´Ó¦”¡£µã»÷»¯Ñ§·´Ó¦×÷ΪһÖÖÖØÒªµÄ·Ö×ÓÁ¬½Ó·½Ê½,Äܹ»ÔÚ·Ö×ÓˮƽʵÏÖÌØ¶¨Ð¡·Ö×Ó»ùÍÅÖ®¼äµÄ¼òµ¥¸ßЧÁ¬½Ó¡£ÒòÆä¿ìËÙ¸ßЧ¡¢Ñ¡ÔñÐԸߡ¢Ìõ¼þκ͡¢¸±·´Ó¦ÉÙµÈÓŵã,ÔÚµ°°×ÖÊ×éѧ·ÖÎöÖÐÓй㷺ӦÓÃ,°üÀ¨ÐÂÉú³Éµ°°×µÄ¼ø¶¨ºÍ¶àÖÖ·­ÒëºóÐÞÊε°°×µÄ¼ø¶¨µÈ·½Ãæ¡£Òò´Ë,ÉîÈëÑо¿µã»÷»¯Ñ§·´Ó¦ÔÚµ°°×ÖÊ×éѧÖеķ´Ó¦»úÖÆ,¶ÔÀí½âµ°°×ÖʵĽṹ¹¦ÄܾßÓÐÖØÒªÒâÒå¡£±¾ÎÄ×ÛÊöÁ˽üÄêÀ´µã»÷»¯Ñ§·´Ó¦ÔÚµ°°×ÖÊ×éѧÖеļ¸ÀàÖØÒªÓ¦ÓÃ,²¢¶ÔÆäδÀ´µÄ·¢Õ¹Ç÷ÊÆ½øÐÐÁËÕ¹Íû¡£

¡¡¡¡¹Ø¼ü´Ê£º¡¡µã»÷»¯Ñ§·´Ó¦; µ°°×ÖÊ×éѧ; ÆÀÊö;

¡¡¡¡Abstract£º¡¡Cu(I)-catalyzed azide-alkyne specific reaction, popularly known as the “click chemistry reaction”, is defined as an important molecular connection method, which can realize simple and efficient connection between specific small molecule groups at the molecular level. Because of its advantages such as high efficiency, high selectivity, mild conditions and few side reactions, click chemistry reaction has been widely used in proteomics analysis, including the identification of newly synthesized proteins and the identification of various post-translational modified proteins. Therefore, in-depth study of the reaction mechanism of click chemistry reaction in proteomics is very important for understanding the structure and function of proteins. The goal of this review is to highlight the progress of several important applications of click chemistry in proteomics analysis in recent years, and discuss the future trends of the click chemistry in proteomics.

¡¡¡¡Keyword£º¡¡Click chemistry reaction; Proteomics; Review;

¡¡¡¡Fund£º supported by the National Nature Science Foundation of China(No.81472030); the Jilin Province Science and Technology Department Project(No.20180101267JC); the Graduate Innovation Fund of Jilin University(No.101832018C070);

¡¡¡¡1 ¡¢Òý ÑÔ

¡¡¡¡µ°°×ÖÊ×é(Proteome)Ô´ÓÚµ°°×ÖÊ(Protein)Óë»ùÒò×é(Genome)Á½¸ö´ÊµÄ×éºÏ,Õâ¸ö¸ÅÄî×îÔçÊÇÓɰĴóÀûÑÇMacquarie´óѧWilkinsµÈ[1]Ìá³ö¡£µ°°×ÖÊ×éѧÑо¿µÄÖ÷ÒªÄÚÈݰüÀ¨ÔÚµ°°×ÖÊˮƽÉÏ´ó¹æÄ£µØ·ÖÎö×é֯ϸ°ûµÄµ°°×Öʱí´ïˮƽ¡¢·­ÒëºóÐÞÊΡ¢µ°°×ÖʼäµÄÏ໥×÷ÓõÈ,´Ó¶ø½Òʾµ°°×ÖʵŦÄÜ,ÒÑÔÚÒßÃçɸѡ¡¢Ö¸µ¼ÖÎÁÆ¡¢ÁÙ´²Ò©Î↑·¢¼°Ô¤ºóÅжϵÈÁìÓò·¢»ÓÁËÖØÒª×÷ÓÃ[2,3]¡£
 

ÐÂÉú³Éµ°°×ºÍÐÞÊε°°×µÄ¼ì²â¼¼Êõ×ÛÊö
 

¡¡¡¡½üÄêÀ´,»¯Ñ§µ°°×ÖÊ×éѧÔÚ»¯Ñ§ÉúÎïѧÁìÓò·¢Õ¹Ñ¸ËÙ,ÆäÀûÓúϳɻ¯Ñ§·½·¨Éú³ÉС·Ö×Ó̽Õë,½Òʾµ°°×ÖʵĽṹºÍ¹¦ÄÜ¡£´«Í³µÄµ°°×±ê¼Ç̽ÕëÖ÷ÒªÓÐÉúÎïËØ¡¢Ó«¹â»ùÍŵÈ,µ«ÆÕ±é´æÔÚ½µµÍµ°°×ÖÊ»îÐÔ¡¢¶Ô¼«ÐԱ仯²»¹»Ãô¸ÐµÈȱµã[4,5]¡£µã»÷»¯Ñ§µÄ³öÏÖ¼«´óµØ½µµÍÁËÒò̽Õë²ôÈë¶Ôµ°°×»îÐÔµÄÓ°Ïì¡£µã»÷»¯Ñ§ÊÇָͨ¹ýС·Ö×Ó»ùÍŵį´½Ó,¼òµ¥¸ßЧµØÍê³ÉC-X-CÔÓÔ­×Ó»¯Ñ§ºÏ³ÉµÄÒ»Àà×éºÏ»¯Ñ§·½·¨¡£Ä¿Ç°, ³£Óõĵã»÷»¯Ñ§·´Ó¦Ö÷Òª°üÀ¨Í­´ß»¯µÄµþµª-Ȳ»ù»·¼Ó³É·´Ó¦(Copper-catalysed azide-alkyne cycloaddi-tion reaction, CuAAC)¡¢Ö±½Ó´Ù½øµÄµþµª-Ȳ»ù»·¼Ó³É·´Ó¦(Strain-promoted azide-alkyne cycloaddition, SPAAC)¡¢Äæµç×ÓÐèÇóµÄ Diels-Alder·´Ó¦(Inverse electron demand Diels-Alder reaction, IEDDA)ºÍÊ©ÌÕ¶¡¸ñÁ´½Ó·´Ó¦(Staudinger ligation)4Àà[6]¡£ÆäÖÐ, CuAACÊǽÏΪ¾­µäµÄµã»÷»¯Ñ§·´Ó¦, ¸Ã·´Ó¦ÊÇÖ¸ÓÉCu+´ß»¯µþµª»¯ºÏÎïÓëȲ»ù»¯ºÏÎï, Éú³ÉÈýßòÎåÔª»·»¯ºÏÎïµÄ¹²¼Û¼Ó³É·´Ó¦, Èçͼ1Ëùʾ¡£¸Ã·½·¨·´Ó¦Ð§Âʸß, ¸±·´Ó¦ÉÙ, ·´Ó¦Ìõ¼þκÍ,¾ßÓкܸߵĻ¯Ñ§Ñ¡ÔñÐÔÇÒ²»ÊÜË®Ñõ¸ÉÈÅ¡¢»·¾³ÎÛȾСµÈÓŵã[6]¡£½üÄêÀ´, µã»÷»¯Ñ§·´Ó¦Óë»ùÓÚÖÊÆ×·¨µÄµ°°×ÖÊ×éѧÑо¿·½·¨µÄ½áºÏÔÚÉúÎïѧÁìÓò¹ã·ºÓ¦ÓÃ, ÌØ±ðÊÇΪÐÂÉú³Éµ°°×[7]¡¢×Øéµõ£»¯ÐÞÊε°°×[8]¡¢ÌÇ»ù»¯ÐÞÊε°°×[9]¼°Á×ËữÐÞÊε°°×µÄ¼ì²âÌṩÁË·½·¨, ÔÚµ°°×ÖÊÓëµ°°×ÖʵÄÏ໥×÷ÓÃ[10]¡¢microRNAÉúÎïºÏ³É[11]ºÍmicroRNAǰÌåÓëµ°°×ÖÊÏ໥×÷ÓÃ[12]·½ÃæÒ²Óй㷺ӦÓᣵã»÷»¯Ñ§µÄʹÓöÔ̽Ë÷µ°°×ÖÊÉúÎïѧÆðµ½Á˾޴óµÄÍÆ¶¯×÷Óᣱ¾ÎĶԻùÓÚµã»÷»¯Ñ§·´Ó¦ÔÚµ°°×ÖÊ×éѧÑо¿Öеļ¸ÀàÖØÒªÉúÎïѧӦÓýøÕ¹½øÐÐÁË×ÛÊö¡£

¡¡¡¡Í¼1 µã»÷»¯Ñ§·´Ó¦Ô­Àí[6]
ͼ1 µã»÷»¯Ñ§·´Ó¦Ô­Àí[6]

¡¡¡¡Fig.1 Schematic of click chemistry reaction[6]

¡¡¡¡2 ¡¢ÐÂÉú³Éµ°°×µÄ¼ì²â

¡¡¡¡µ°°×ÖʵĺϳÉÊÇÒ»¸ö¶¯Ì¬¹ý³Ì, ÔÚÕý³£ºÍ²¡ÀíÌõ¼þÏÂ, ϸ°û»áͨ¹ý²úÉúÐÂÉú³Éµ°°×¶Ô»·¾³±ä»¯×ö³ö·´Ó¦, ׼ȷ¼ì²âµ°°×ÖʶԻ·¾³ÈŶ¯Ëù×ö³öµÄ·´Ó¦¶ÔÀí½âÆäDZÔÚ×÷ÓûúÖÆ¾ßÓÐÊ®·ÖÖØÒªµÄÒâÒ塣оɵ°°×Öʹ²ÏíÏàͬµÄ°±»ùËá¿â, ÔÚÉúÎïѧÉÏÄÑÒÔÇø·Ö, Òò´ËÓбØÒª¶ÔÐÂÉú³Éµ°°×½øÐÐÌØÊâ±ê¼ÇÒÔ½øÐж¨ÐÔÓ붨Á¿·ÖÎö¡£´«Í³±ê¼Ç·½·¨Ê¹ÓÃÎȶ¨Í¬Î»Ëذ±»ùËá±ê¼Ç·¨(Stable isotope labelling by amino acids in cell culture, SILAC)[13]±È½ÏÈŶ¯Ç°ºóµ°°×Öʵıí´ï±ä»¯¡£Ïà½ÏÓÚ¼«¸ß·á¶ÈµÄÔ­Óе°°×ÖÊ, ÐÂÉú³Éµ°°×Öʳ£Òò·á¶ÈÌ«µÍ, ÎÞ·¨ÊµÏÖÔÚ´ó¶àÊýÖÊÁ¿·ÖÎöÒǵ͍Á¿¶¯Ì¬·¶Î§Äڵļì²â[14], Òò´ËSILAC¶ÔÓÚ²âÁ¿µ°°×ÖʺϳɵĶÌʱ¶ÎÄڱ仯²¢²»ÊÇ×î¼ÑµÄ·½·¨¡£ÒÑÓÐÑо¿·¢ÏÖÁËÁ½ÖÖ·ÇÌìÈ»°±»ùËáµþµª¸ß±û°±Ëá(Azidohomoalanine, AHA)ºÍ¸ßȲ±û»ù¸Ê°±Ëá(Homopropargylglycine, HPG), ËüÃÇ¿ÉÒÔÔÚϸ°ûµÄÕý³£µ°°×ÖʺϳÉʱ²ôÈëµ½¸Ãµ°°×µÄÒ»¼¶½á¹¹ÖÐ, ²¢ÇÒÎÞÃ÷ÏÔϸ°û¶¾ÐÔ[15,16]¡£ÉúÎïÕý½»·ÇÌìÈ»°±»ùËá±ê¼Ç·¨(Bioorthogonal noncanonical amino acid tagging, BONCAT, Èçͼ2Ëùʾ[17])ÀûÓôËÌØµã, ͨ¹ý´úл±ê¼ÇÒýÈëAHA»òHPG[18,19]ÕâÖÖС·Ö×Ó»ùÍż×Áò°±ËáÀàËÆÎï¡£½«±»±ê¼ÇµÄϸ°û½øÐÐÁѽâºó, ͨ¹ýµã»÷»¯Ñ§·´Ó¦½«´øÓÐȲ»òµþµª»¯ÎïµÄÇ׺ͱêÇ©¹²¼ÛÁ¬½ÓÔÚÄ¿µÄµ°°×ÉÏ, ÔÙ¾­¹ýÇ׺ÍËØÇíÖ¬ÌÇÄýÖé¶ÔÄ¿µÄµ°°×½øÐд¿»¯¸»¼¯, ×îºó½«¸»¼¯²úÎïø½â³É¶àëÄ, ÀûÓøßͨÁ¿ÖÊÆ×½øÐзÖÎö¼ì²â¡£¸Ã·½·¨¾ßÓÐÌØÒìÐÔÇ¿¡¢ ÁéÃô¶È¸ßµÈÓŵã[20,21], ¿ÉÒÔÑ¡ÔñÐÔ¸»¼¯ÐÂÉú³Éµ°°×, ʵÏÖ¶ÔÐÂÉú³Éµ°°×¸üÉîÈëµÄ·ÖÎö, ´Ó¶ø´Ù½øÏ¸°û¼äÏ໥×÷ÓõÄÉúÎïѧµÄÑо¿·¢Õ¹¡£

¡¡¡¡Í¼2 ʹÓÃBONCAT·½·¨¶ÔÐÂÉú³Éµ°°×½øÐбê¼Ç¡¢¼ì²âºÍ¼ø¶¨[17]¡¢
ͼ2 ʹÓÃBONCAT·½·¨¶ÔÐÂÉú³Éµ°°×½øÐбê¼Ç¡¢¼ì²âºÍ¼ø¶¨[17]¡¢

¡¡¡¡Fig.2 Bioorthogonal noncanonical amino acid tagging (BONCAT) strategy for labeling, detection and identification of newly synthesized proteins[17]

¡¡¡¡¸Ã·½·¨×Ô½¨Á¢ÒÔÀ´, ÔÚ¶¯Ì¬µ°°×ÖÊ×éѧµÄ·ÖÎöÖеõ½¹ã·ºÓ¦Óá£ShenµÈ[22]²ÉÓÃBioorthogonal noncanonical amino acid tagging (BONCAT)ÓëMSÁªÓü¼Êõ¶Ô²¸È鶯Îïϸ°û½øÐÐÁËÑо¿, ·¢ÏÖÁËÊý°Ù¸öÐÂÉú³Éµ°°×ÔÚÊÓ¾õÌõ¼þʱÊÓÉñ¾­Ö§ÅäÇø¼±¾çÔö¼Ó; ×÷Õß½øÒ»²½·¢ÏÖ, Rab3¼Ò×å³ÉÔ±ÖеÄÍ»´¥µ°°×Rab3a¡¢ Rab3bºÍRab3cÊܵ½ÊÓ¾õÉñ¾­µÄ²îÒìµ÷½Ú, ÔÚÊÓ¾õ´Ì¼¤µÄ¶¥¸ÇÄÔÖÐRab3bºÍRab3cµÄ±í´ïÁ¿ÏÔ×ÅÔö¼Ó, ¶øRab3aÏÔ׿õÉÙ, ˵Ã÷Rab3¼Ò×å³ÉÔ±¿ÉÄÜÔÚÌåÄڽ鵼ÊÓ¾õÒÀÀµµÄÐÐΪ¿ÉËÜÐÔÖаçÑݲ»Í¬µÄ½ÇÉ«¡£HowdenµÈ[23]ÀûÓÃAHA±ê¼Ç²¢½áºÏÁËSILACµÄ·½·¨(Quantitative non-canonical amino acid tagging, QuaNCAT), Ö»ÐèÒª±ê¼Çϸ°ûÊýСʱ»òιʳ/×¢È붯Îï1¡«4Ìì, ¼´¿ÉʵÏֶԵͷá¶ÈÐÂÉú³Éµ°°×µÄ¶¨Á¿·ÖÎö¡£ÔÚ¸ÃÑо¿ÖÐ, ÀûÓÃÖØÍ¬Î»ËØ±ê¼ÇµÄ°±»ùËá¶ÔTϸ°û´Ì¼¤ºó·ÖÃÚµÄÐÂÉú³Éµ°°×½øÐбê¼Ç, Ëæºóͨ¹ýÇ׺ÍËØÎ¢Öé¶ÔÄ¿µÄµ°°×½øÐи»¼¯, ×îºóÀûÓÃMS¶ÔÐÂÉú³Éµ°°×½øÐÐÁ˶¨Á¿¼ì²â¡£MaµÈ[24]µÄ·½·¨¸ü¼òµ¥Ö±½Ó, ʹÓÃÖØÍ¬Î»ËØ±ê¼ÇµÄAHA(Heavy isotope-labeled azidohomoalanine quantification, HILAQ)½øÐÐÐÂÉú³Éµ°°×µÄ±ê¼Ç, ÔÙͨ¹ýµã»÷·´Ó¦Ìí¼ÓÉúÎïËØ, ½Ó×ŶԱê¼ÇµÄµ°°×½øÐгÁµíÏ´µÓºÍÏû»¯¡£¸ÃÑо¿×é¶ÔHT22ϸ°ûÑõ»¯Ä£Ð͵ÄÐÂÉú³Éµ°°×½øÐÐÁ˶¨Á¿¼ì²â, ·¢ÏÖ226ÖÖµ°°×Öʵķá¶È·¢ÉúÁËÁ½±¶µÄ±ä»¯, 108ÖÖµ°°×ÖÊÔÚϸ°ûËÀÍö;¾¶Öеõ½¸»¼¯, Ö¤Ã÷ÁËHT22ϸ°û×÷ΪһÖÖ·Ö×Ó¹¤¾ß, ÔÚÑо¿Éñ¾­ÍËÐÐÐÔ¼²²¡Éæ¼°µÄϸ°ûµòÍöµÄ¹ý³ÌÖÐÊ®·ÖÓÐЧ¡£ËûÃÇ»¹½«HILAQ·¨ÓëQuaNCAT·¨½øÐбȽϷ¢ÏÖ, HILAQʹÓÃëÄˮƽ¸»¼¯, Òѱ»Ö¤Ã÷ÊDZÈQuaNCATÖÐʹÓõ°°×ÖÊˮƽ¸»¼¯µÄ¸üÁéÃôµÄ·½·¨, ÇÒHILAQ·¨¶¨Á¿µÄÐÂÉú³Éµ°°×ÊÇQuaNCAT·¨¶¨Á¿µÄ3.5±¶ÒÔÉÏ¡£×ÛÉÏ, HILAQ·½·¨ÊÇÒ»ÖÖÐÂÐ͵ÄÐÂÉú³Éµ°°×µÄ¶¨Á¿¼ì²â·½·¨, ͨ¹ýʹÓÃÖØÍ¬Î»ËØµÄ±ê¼Ç¼ò»¯ÁËкϳɵ°°×ÖÊ×éµÄ·ÖÎö¹ý³Ì, ²¢ÇÒ¾ßÓиü¸ßµÄÁéÃô¶È¡£´ÞÐãÔÆµÈ[25]Ê×ÏÈÀûÓÃÖ¬¶àÌǴ̼¤Raw264.7ϸ°û²úÉúÖ×Áö»µËÀÒò×Óα, È»ºóͨ¹ý´úл²ôÈëAHA, ²¢ÀûÓõã»÷»¯Ñ§·´Ó¦, ʹϸ°ûÖеÄÐÂÉú³ÉµÄµ°°×±ê¼ÇÉÏÉúÎïËØ±êÇ©, ÔÙÀûÓÿ¹Ìå¶Ô±ê¼Çµ°°×½øÐÐÌØÒìÐÔ²¶»ñ, ´Ó¶ø´ïµ½¼ì²âÌØ¶¨µÄÐÂÉú³Éµ°°×µÄÄ¿µÄ¡£¸ÃÑо¿Îª¼ì²âÌØ¶¨µÄ΢Á¿ÐÂÉú³Éµ°°×ÌṩÁËÒ»ÖÖ¿É½è¼øµÄ·½·¨¡£

¡¡¡¡³ýÁËÉÏÊöAHA»òHPG, Ò»ÖÖ·ÇÌìÈ»°±»ùËá¶Ô-µþµª-L-±½±û°±Ëá(p-azido-L-phenylalanine, azF)Ò²±»ÓÃÓÚµ°°×ÖʵĴúл±ê¼Ç[26,27]¡£SmitsµÈ[28]¿ª·¢ÁËÒ»ÖÖClick-MSµÄÐÂÐͼì²â·½·¨, ÀûÓÃçúçêÒÖÖÆ¼¼ÊõÑ¡ÔñÐԵؽ«çúçêÖÕÖ¹ÃÜÂë×Ó(UAG)²ôÈ뵽ĿµÄµ°°×ÖÐ, ͬʱÌí¼ÓÌØ¶¨µÄtRNAºÍõ£°±»ù-tRNAºÏ³Éø(aa-RS)¡£È»ºó, ͨ¹ýÎÞÍ­µã»÷»¯Ñ§·´Ó¦¶Ô²ôÈë¶Ô-µþµª-L-±½±û°±ËáµÄÄ¿µÄµ°°×½øÐÐÌØÒìÐÔ²¶»ñ, ×îºóÀûÓÃÖÊÆ×¼ì²â, ʵÏÖµ¥¸öÄ¿µÄµ°°×µÄÑ¡ÔñÐÔ¸»¼¯¡£Ñо¿ÕßÀûÓô˷½·¨, ´Ó²¸È鶯Îïϸ°û´ÖÌáÈ¡ÎïÖÐÌØÒìÐÔ¸»¼¯ÁËÁ½ÖÖÄ¿±êµ°°×: ÐźÅת¼Ó뼤»îÒò×Ó1(STAT1)ºÍ¼×»ùCpG½áºÏÓòµ°°×3(MBD3)¡£ÓëʹÓÃAHA»òHPGµÄµ°°×Öʱê¼Ç²ßÂÔÓÐËù²»Í¬, ¸Ã·½·¨ÊµÏÖÁ˶Ե¥¸öÄ¿µÄµ°°×µÄÌØÒìÐÔ¸»¼¯, ¶ÔÓÚ½ÒÊ¾ÌØ¶¨Ä¿±êµ°°×µÄ´úлÇé¿öÒÔ¼°ÓëÆäËüµ°°×¼äµÄÏ໥×÷ÓþßÓÐÖØÒªÒâÒå¡£

¡¡¡¡3¡¢ ·­ÒëºóÐÞÊε°°×µÄ¼ì²â

¡¡¡¡3.1¡¢ רéµõ£»¯ÐÞÊε°°×µÄ¼ì²â

¡¡¡¡×Øéµõ£»¯ÐÞÊε°°×µÄ´«Í³¼ì²â·½·¨ÊÇõ£»ùÉúÎïËØÖû»·¨ (Acyl-biotinyl exchange, ABE)[29]¡£ ABE·¨ÊÇÏÈÓõâÒÒõ£°··â±Õµ°°×Öʰëë×°±ËáÉϵÄ×ÔÓÉÛÏ»ù, ÀûÓÃôǰ·Ñ¡ÔñÐÔµØÇи°×Öʰëë×°±ËáºÍÖ¬·¾ËáÐÞÊλùÍÅÖ®¼äµÄÁòõ¥¼ü, ÔÙ½«´øÓÐÉúÎïËØµÄÛÏ»ùÌØÒìÐÔ·´Ó¦»îÐÔÊÔ¼ÁÓëвúÉúµÄ×ÔÓÉÛÏ»ù·´Ó¦, ×îºó¶ÔÄ¿µÄµ°°×S-רéµõ£»¯ÐÞÊε°°×½øÐд¿»¯¡¢¸»¼¯ºÍ¼ì²â¡£ÒÑÓÐÑо¿²ÉÓÃABE·¨¶ÔÉúÎïÑù±¾ÖеÄרéµõ£»¯ÐÞÊε°°×½øÐÐÁËÊýÁ¿¼°ÐÞÊÎλµãµÄ¼ø¶¨·ÖÎö[30,31]¡£¾¡¹ÜABE·¨ÔÚ¼ì²âרéµõ£»¯ÐÞÊε°°×ÖÊ·½ÃæÓÐÁ˺ܴó½øÕ¹, µ«ÓÉÓÚôǰ·ÇиîµÄÊÇÁòõ¥¼ü, ¶ø²»ÊÇS-רéµõ£»¯ÐÞÊεÄÌØÒìλµã, ÆäËü¶Ôôǰ·Ãô¸ÐµÄÁòõ¥¼üµ°°×Ò²ÓпÉÄܱ»Í¬Ê±¸»¼¯[32], Òò´Ë´æÔÚ¼ÙÑôÐÔÂʸߡ¢ÁéÃô¶ÈµÍ¡¢·ÅÉäÐÔÇ¿µÈȱµã, ¼«´óÏÞÖÆÁËÔÚרéµõ£»¯ÐÞÊε°°×¼ì²â·½ÃæµÄÓ¦Óá£

¡¡¡¡½üÄêÀ´, Ñо¿Õß·¢Õ¹ÁËÁíÒ»ÖÖµ°°×ÖÊרéµõ£»¯ÐÞÊεķÖÎö·½·¨: »¯Ñ§±¨¸æ»ùÍÅ·¨(Chemical Reporter)[33], ¸Ã·½·¨Äܹ»Çø·ÖS-Ö¬·¾õ£»¯(רéµõ£»¯ÐÞÊÎ)µÄµ°°×ºÍN/O-Ö¬·¾õ£»¯µÄµ°°×¡£ÓëÉÏÊöABE·¨²»Í¬µÄÊÇ, ¸Ã·½·¨½«º¬ÓÐȲ»ùµÄרéµËáÀàËÆÎï´úл²ôÈëϸ°ûµ°°×ÖÊ, È»ºóͨ¹ýµã»÷»¯Ñ§·½·¨, Ñ¡ÔñÐԵؽ«´øµþµª»ùÍŵÄÉúÎïËØÁ¬½Óµ½±ê¼ÇÁËȲ»ù̽ÕëµÄµ°°×ÖÊÉÏ, ½Ó×ÅʹÓÃôǰ·¶Ôרéµõ£»¯ÐÞÊε°°×µÄÁòõ¥¼ü´¦½øÐÐÌØÒìÐÔÇиî, ×îºó²ÉÓÿ¹ÉúÎïËØÇíÖ¬ÌÇÄýÖé¶Ôµ°°×ÖʽøÐи»¼¯ºÍ¼ø¶¨, ´Ó¶øÊµÏÖ¶Ôרéµõ£»¯ÐÞÊε°°×µÄ¼ì²â, Èçͼ3Ëùʾ¡£È²ÌþºÍµþµª»¯ÎïÖ¬·¾Ëá̽Õëͨ³£¶¼ÄÜÓÐЧ±ê¼Çϸ°ûÖ¬·¾õ£»¯µ°°×ÖÊ, ÆäÖÐȲ»ù̽ÕëÒò¾ßÓнϵ͵ı³¾°Ðźż°¸ü¸ßµÄ¼ì²âÁéÃô¶ÈºÍ¼ì²âЧÂʵÈÓŵãÓ¦ÓøüΪ¹ã·º[34]¡£Í¬Ê±, Ìí¼ÓÒ»¸öµþµª»ùÍŵ½Ö¬·¾ËáÁ´ÉÏ¿ÉÄÜ»áÓ°ÏìÖ¬·¾ËáµÄÊèË®ÐÔ, ¶øÈ²»ù¿É±£ÁôÖ¬·¾Ëá̼Á´µÄÊèË®ÐÔ, ¼õÉÙ¶ÔÖ¬·¾ËáÎïÀí»¯Ñ§ÐÔÖʵĸı估ÆäÓëÖ¬ÖʼäµÄÏ໥×÷ÓõÄÓ°Ïì[32]¡£YountµÈ[35]ÔÚÍÌÊÉϸ°û(ÊóÊ÷ͻ״ϸ°ûϵ)ÖÐʹÓÃAlk-C16½øÐдó¹æÄ£×Øéµõ£»¯Ã¸Í¼Æ×·ÖÎö, ¼ø¶¨³ö157ÖÖ¾ßÓжàÖÖ¹¦ÄÜ, °üÀ¨ÏÈÌìÃâÒßµÄS-רéµõ£»¯µ°°×(ÆäÖÐ60¸ö¸ß¿ÉÐŶȵ°°×ºÍ97¸öÖпÉÐŶȵ°°×)¡£ËûÃÇÑо¿·¢ÏÖרéµõ£»¯¶ÔÓÚ¸ÉÈÅËØÓÕµ¼µÄ¿çĤµ°°×(IFITM3)µÄ¿¹²¡¶¾»îÐÔÖÁ¹ØÖØÒª, ¸Ãµ°°×ÔÚÆäĤÉÏ3¸ö½ü°ëë×°±Ëá²Ð»ù(Cys71/72/105)´¦±»×Øéµõ£»¯¡£´ËÍâ, TeoµÈ[36]½«»ùÓÚµã»÷»¯Ñ§µÄ̽ÕëÓÃÓÚ·ÖÎö¸ÐȾµ¥´¿ðåÕ¶¾(HSV)µÄËÞÖ÷ϸ°ûÖеÄÕûÌåÖ¬·¾õ£»¯, ·¢ÏÖËÞÖ÷Sרéµõ£»¯µ°°×µÄÒ»¸ö×Ó¼¯(°üÀ¨¸ÉÈÅËØºÍËÄ¿çĤµ°°×¼Ò×å³ÉÔ±)±»Ñ¡ÔñÐÔÒÖÖÆ, ÔÚ¸ÐȾ¹ý³ÌÖÐ, ºÜ´óÒ»²¿·ÖHSVµ°°×¾ù±»S-רéµõ£»¯¡£

¡¡¡¡Í¼3 ²ÉÓû¯Ñ§±ê¼Ç·¨¼ø¶¨×Øéµõ£»¯ÐÞÊε°°×[33]
ͼ3 ²ÉÓû¯Ñ§±ê¼Ç·¨¼ø¶¨×Øéµõ£»¯ÐÞÊε°°×[33]

¡¡¡¡Fig.3 Identification of palmitoylated proteins by chemical labeling[33]

¡¡¡¡3.2 ¡¢ÌÇ»ù»¯ÐÞÊε°°×µÄ¼ì²â

¡¡¡¡µ°°×ÖʵÄÌÇ»ù»¯ÐÞÊÎÊÇÒ»ÖÖÖØÒªÇÒ³£¼ûµÄ·­ÒëºóÐÞÊÎ, Õ¼ÈËÀ൰°×ÖÊ×ÜÁ¿µÄÒ»°ëÒÔÉÏ, ¸ÃÐÞÊη½Ê½²ÎÓëÐí¶àÉúÀí¹¦ÄܺÍÉúÎïѧ;¾¶[37]¡£ÔÚÐí¶à¼²²¡Öж¼Äܹ۲쵽ÌÇ»ù»¯ÐÞÊε°°×µÄÒì³£, Èç×ÔÉíÃâÒßÐÔ¼²²¡¡¢ÐÄѪ¹Ü¼²²¡¡¢°¢¶û×Ⱥ£Ä¬Ö¢ºÍ°©Ö¢µÈ[38,39], Òò´Ë¶ÔÉúÀí²¡ÀíÌõ¼þϵÄÌÇ»ù»¯ÐÞÊε°°×½øÐж¯Ì¬¼à²â¶ÔÓÚÀí½â¼²²¡µÄ·¢Éú·¢Õ¹»úÖÆ¾ßÓÐÖØÒªÒâÒå¡£

¡¡¡¡µã»÷»¯Ñ§½éµ¼µÄÌÇ»ù»¯µ°°×µÄ±ê¼Ç(Glycoprotein labeling with click chemistry, GLCC)ÊÇÒ»ÖÖ³£Óüì²âÊÖ¶Î[40]¡£ÔÚÕâÖÖ·½·¨ÖÐ, º¬ÓÐÉúÎïÕý½»¹¦ÄÜ»ùÍÅ(Èçµþµª)µÄµ¥ÌÇÀàËÆÎïͨ¹ýϸ°û×ÔÉíµÄ´úл»úÖÆ²ôÈëϸ°ûµÄ¾ÛÌÇÖÐ, ÔÙÀûÓõã»÷»¯Ñ§·´Ó¦½«´øÓÐÉúÎïËØ»òÓ«¹â»ùÍŵÄȲ»ù±ê¼Ç·Ö×ÓÁ¬½Óµ½ÌÇ»ù»¯µ°°×ÉÏ, ×îºóͨ¹ýÁ´Ã¹Ç׺ÍËØÇíÖ¬ÌÇÄýÖé»òÓ«¹â¹²¾Û½¹ÏÔ΢¾µ³ÉÏñ¶Ô±ê¼Ç·Ö×Ó½øÐи»¼¯ºÍ¼ì²â¡£Ä¿Ç°, ÒÑÓдóÁ¿Ñо¿³É¹¦ÀûÓõã»÷»¯Ñ§·´Ó¦¶ÔÌÇ»ù»¯µ°°×ʵÏÖÁË¿ÉÊÓ»¯µÄ¶¯Ì¬¼ì²â¡£RongµÈ[41]½«µþµª×÷Ϊ»¯Ñ§±¨¸æ»ùÍÅ, ´úл²ôÈëµ½´óÊóÐÄÔà×éÖ¯ÖÐ, ÀûÓõã»÷»¯Ñ§·´Ó¦ÔÚµþµªÉÏÁ¬½Ó´øÓ«¹â»ùÍŵÄȲ»ù·´Ó¦ÊÔ¼Á, ×îºóͨ¹ýÓ«¹â¹²¾Û½¹ÏÔ΢¾µ³ÉÏñ¼ø¶¨³ö³¬¹ý200ÖÖÍÙÒºËáÐÞÊεÄÐ°°×ÈçT-kininogensµÈ, ²¢¶ÔÍÙÒºËữµÄ¾ÛÌǺÍOÁ¬½ÓµÄ¾ÛÌǽøÐÐÁ˸߷ֱæÂʵijÉÏñ·ÖÎö, ·¢ÏÖËüÃÇÖ÷Òª·Ö²¼ÔÚϸ°ûÓëϸ°û¼äµÄÁ¬½Ó´¦ºÍTÐ͹ܵÄϸ°ûĤÉÏ¡£¸ÃÑо¿»¹Ö¤Ã÷, µã»÷»¯Ñ§·´Ó¦½éµ¼µÄÌǵ°°×±ê¼ÇÔÚϸ°ûˮƽ¡¢¶ÀÁ¢µÄÐÄÔàÆ÷¹ÙˮƽºÍ¶àëÄˮƽ¶¼ÄÜʵÏÖ±ê¼Ç, ²¢³É¹¦½øÐмì²â(ͼ4)¡£ÆäÖÐ, ÔÚϸ°ûˮƽ²ÉÓÃÒ»¼ÛÍ­Àë×Ó´ß»¯µÄµã»÷·´Ó¦½«´úл²ôÈëµÄµþµªÌÇÓëȲ»ùÓ«¹â488·Ö×ÓÏàÁ¬½Ó, ͨ¹ýÓ«¹â¹²¾Û½¹ÏÔ΢¾µ³ÉÏñ½øÐмì²â; ÔÚ¶ÀÁ¢ÐÄÔàÆ÷¹Ùˮƽ, ¶Ô±ê¼ÇÁ˵þµªÌǵÄÐÄÔà×éÖ¯½øÐÐLangendorff¹à×¢, ¿¼Âǵ½Í­Àë×ÓµÄÉúÎï¶¾ÐÔ, Ñо¿²ÉÓõÄÊDz»ÒÀÀµÓÚÍ­µÄDBCO-Ó«¹â488»ùÍűê¼Ç´úл²ôÈëµÄµþµªÌÇ, ²¢¶ÔÍêÕûµÄÐÄÔà½øÐÐÓ«¹â³ÉÏñ·ÖÎö; ÔÚ×éÖ¯ÁѽâҺˮƽ, ½«²ôÈëÁ˵þµª»¯ÎïµÄÐÄÔà×éÖ¯½øÐÐÁѽâºó, ÓëȲ»ùÉúÎïËØ½øÐеã»÷·´Ó¦, ÓÃÁ´Ã¹Ç׺ÍËØ¸»¼¯ÍÙÒºËữµÄÌǵ°°×, È»ºó½øÐÐÄý½ºË®Æ½µÄµ°°×¼ø¶¨¡£ÕâЩʵÑé¶¼Ö¤Ã÷ÁË´óÊóÐÄÔàÌÇÔ­µÄÌåÄÚ´úл±ê¼Ç¿ÉʹÍêÕûÐÄÔà×éÖ¯ÖеÄÐļ¡Ï¸°û±íÃæ¾ÛÌÇ¿ÉÊÓ»¯, ²¢·¢ÏÖÔÚ¶ÀÁ¢Æ÷¹ÙˮƽµÄÐļ¡·Ê´óÄ£ÐÍСÊóµÄ²¡Àí¹ý³ÌÖÐ, ÍÙÒºËữ¾ÛÌǵÄÉúÎïºÏ³ÉÉϵ÷, Ö¤Ã÷ÍÙÒºËữ¶ÔÐÄÔà·¢ÓýºÍ·¢²¡»úÖÆ´æÔÚDZÔÚÓ°Ïì¡£HerberµÈ[42]²ÉÓõã»÷·´Ó¦¸»¼¯ÌDZíÃæµ°°×(Surface-spanning protein enrichment with click sugars, SUSPECS)·½·¨, ʵÏÖÁËϸ°û±íÃæÌǵ°°×µÄ¸»¼¯¡¢¼ø¶¨ºÍ¶¨Á¿·ÖÎö¡£¸ÃÑо¿ÏÈʹÓõã»÷»¯Ñ§·´Ó¦Ñ¡ÔñÐÔ±ê¼Çϸ°û±íÃæÌÇ»ù»¯ÐÞÊε°°×, ¶øºóͨ¹ý¶ÔÉúÎïËØµÄ¼ì²â, ¶ÔÌÇ»ù»¯ÐÞÊε°°×½øÐиßͨÁ¿ÖÊÆ×¶¨Á¿·ÖÎö¡£»ùÓÚÃâÒßÓ«¹â¹²¾Û½¹ÏÔ΢¾µÏÔʾ, SUSPECSÑ¡ÔñÐÔ±ê¼ÇÁËϸ°û±íÃæµ°°×, ÔÚÔ­´úÉñ¾­Ôª±íÃæ´æÔÚ½ü700ÖÖ¿çĤÌǵ°°×¡£ËûÃÇ»¹½«¸Ã·½·¨Ó¦ÓÃÓÚÖÎÁư¢¶û×Ⱥ£Ä¬²¡µÄ¹Ø¼üÒ©Îï°Ð±êµ°°×øBACE1µÄÑо¿ÖÐ, ·¢ÏÖBACE1ͨ¹ýÒÖÖÆÃ¸´úл, Ñ¡ÔñÐÔÖØËÜÁËÉñ¾­Ôª±íÃæÌǵ°°××é, ÕâÖÖ½«µã»÷»¯Ñ§·´Ó¦Óë¼²²¡¹Ø¼üÒ©Îï°Ð±êµ°°×¼ì²âÏà½áºÏµÄÊֶζԽÒʾ°¢¶û×Ⱥ£Ä¬²¡µÄ·¢²¡»úÖÆ¾ßÓÐÖØÒªÒâÒå¡£SongµÈ[43]½«º¬µþµªµÄAC4ManNAZ´úл²ôÈ뵽ϸ°ûĤ±íÃæ, ÀûÓÃÍ­´ß»¯µÄµã»÷»¯Ñ§·´Ó¦¸»¼¯ÌÇ»ù»¯ÐÞÊε°°×, ͨ¹ýÕâÖÖ·½Ê½Äܹ»È¥³ý·á¶È½Ï¸ßµÄ°ûÖʵ°°×, ¸üºÃµØ±íÕ÷ϸ°ûĤ±íÃæµÄÌÇ»ù»¯ÐÞÊε°°×¡£YangµÈ[44]³É¹¦¶ÔǰÁÐÏÙ°©Ï¸°ûϵÖеÄÍÙÒºËữÌǵ°°×½øÐÐÁ˼ø¶¨¡£ÔÚ·Ç×ªÒÆÐÔN2ϸ°ûϵÖй۲쵽36ÖÖϸ°û±íÃæµÄÌǵ°°×, ÆäÖÐÓÐ8ÖÖÌǵ°°×²ÎÓëϸ°ûµòÍö¹ý³Ì; ÔÚ×ªÒÆÐÔML2ϸ°ûϵÖз¢ÏÖ44ÖÖϸ°û±íÃæµÄÌǵ°°×, ÆäÖÐÓÐ11ÖÖÌǵ°°×Óëϸ°ûÔ˶¯¡¢Ç¨ÒƺÍÇÖÏ®Óйء£ÔÚÉÏÊöÓëϸ°ûÇÖÏ®ÓйصÄÌǵ°°×ÖÐ, ÓÐ6ÖÖÌǵ°°×²»ÂÛÊÇ×ªÒÆÐÔ»¹ÊÇ·Ç×ªÒÆÐÔ¾ù¿ÉÓÃ×÷ÉúÎï±ê¼Ç¡£¸ÃÑо¿²ßÂÔ¶ÔǰÁÐÏÙ°©·¢Éú·¢Õ¹¹ý³ÌµÄÑо¿Æðµ½Á˼«´óµÄÍÆ¶¯×÷Óá£

¡¡¡¡Í¼4 ͨ¹ýÌåÄÚ´úл±ê¼Ç, ÔÚϸ°ûˮƽ¡¢Æ÷¹ÙˮƽºÍ×éÖ¯ÁѽâҺˮƽ¶ÔСÊóÐÄÔàÍÙÒºËữÌǵ°°×½øÐе°°×ÖÊ×éѧ¼ø¶¨[41]
ͼ4 ͨ¹ýÌåÄÚ´úл±ê¼Ç, ÔÚϸ°ûˮƽ¡¢Æ÷¹ÙˮƽºÍ×éÖ¯ÁѽâҺˮƽ¶ÔСÊóÐÄÔàÍÙÒºËữÌǵ°°×½øÐе°°×ÖÊ×éѧ¼ø¶¨[41]

¡¡¡¡Fig.4 Proteomic identification of mouse cardiacsialylated glycoproteins by in vivo metabolic labeling at the cell level, organ level and tissue lysate level[41]

¡¡¡¡Ä¿Ç°, µã»÷»¯Ñ§·´Ó¦²»¾ÖÏÞÓÚµþµªºÍȲ»ùµÄ·´Ó¦, »¹ÓÐÒ»ÀàÐÂÐ͵ĵã»÷»¯Ñ§·´Ó¦——Áò´¼ºÍÏ©µÄÌØÒìÐÔ·´Ó¦[45], ÔÚ·Ö×ÓÓ¡¼£¼¼Êõ±ê¼ÇÌǵ°°×·½Ã淢չѸËÙ¡£Ê×ÏÈ, ·Ö×ÓÓ¡¼£ÊÇÒ»ÖÖÔÚÄ£°å·Ö×Ó´æÔÚÏÂÖÆ±¸ÌØÒìʶ±ð¾ÛºÏÎïµÄ¼¼Êõ, Òò¾ßÓгɱ¾µÍÇÒÒ×ÓÚÖÆ±¸µÄÓŵã, ±»³É¹¦Ó¦ÓÃÓÚ·Ö×ӵķÖÀëºÍʶ±ð[46]¡£Ïà±ÈÓÚÒÔÍùÓ«¹â¹¦Äܵ¥Ìå·±ËöµÄÉè¼ÆºÏ³É¹ý³Ì, ÐÂÐÍ·Ö×ÓÓ¡¼£Ó«¹â´«¸ÐÆ÷µÄºÏ³ÉÎÞÐè¶îÍâµÄÓ«¹â²ÄÁÏ(Á¿×ӵ㡢̼µãºÍÓ«¹âËØµÈ), Ö»Ðèͨ¹ý¼òµ¥µÄµã»÷·´Ó¦¼´¿É½«ÆÕͨµÄµ°°×ÖÊÓ¡¼£¾ÛºÏÎïת±ä³ÉÒ»ÖÖÐÂÐ͵ķÖ×ÓÓ¡¼£Ó«¹â´«¸Ð²ÄÁÏ, ¿ÉÓ¦ÓÃÓÚÉúÎï´ó·Ö×Ó(Èçµ°°×ÖʺͶàëÄ)µÄʶ±ð¡£ZhaoµÈ[47]ÀûÓøýáºÏ¼¼Êõ, ½«ÆÕͨ·Ö×ÓÓ¡¼£¾ÛºÏÎïÉý¼¶ÎªÉúÎï´«¸ÐÆ÷Ó«¹â´«¸Ð²ÄÁÏ, ³É¹¦ÊµÏÖÁËÌǵ°°×µÄ¿ìËÙ¼ì²â¡£ËûÃÇʹÓú¬ÓжþÁò¼üµÄ¹¦Äܵ¥Ìå, ÔÚ¶þÁò¼ü¶ÏÁѺó, ½«±©Â¶µÄÁò´¼»ùÍÅͨ¹ý“Áò´¼-Ï©”µÄµã»÷·´Ó¦Á¬½ÓÉÏ´øÓÐÓ«¹âÌØÐÔºÍλµãÌØÒìÐÔʶ±ðµÄ¹¦ÄÜ»ùÍÅ¡£Í¨¹ýÒ»²½Î»µãÌØÒìÐÔ·´Ó¦, ²»½öÒýÈëÁËÓëÌǵ°°×¾ßÓÐÇ׺ÍÁ¦µÄ¹ÙÄÜÍÅ, »¹¸³ÓèÁËÆÕͨµ°°×Ó¡¼£¾ÛºÏÎïÁ¼ºÃµÄÓ«¹âÌØÐÔ(ÏßÐÔ·¶Î§Îª0.1¡«10 mg/mL), ʹÆÕͨµ°°×Ó¡¼£¾ÛºÏÎïÖй¦ÄÜ»ùÍŵÄת»»¸ü·½±ã¼òµ¥¡£

¡¡¡¡3.3 ¡¢Á×ËữÐÞÊε°°×µÄ¼ì²â

¡¡¡¡µ°°×ÖʵÄÁ×ËữÐÞÊÎÊÇÒ»ÖÖÖØÒªµÄ·­ÒëºóÐÞÊÎ, ²ÎÓëϸ°ûµÄÔöÖ³·Ö»¯¡¢ÐźÅתµ¼¡¢Ð³´úлµÈÉúÃü»î¶¯¹ý³Ì, ÊÇÉúÎïÌåÄÚ×îÖØÒªµÄµ÷½Ú»úÖÆÖ®Ò»[48]¡£Òì³£µÄÁ×Ëữµ÷½Ú»áµ¼ÖÂÐí¶àÑÏÖØµÄÈËÀ༲²¡[49], ¶ÔÁ×ËữÐÞÊε°°×½øÐмì²â·ÖÎöÓÐÖúÓÚÍÆ½ø¼²²¡·¢²¡»úÖÆµÄÑо¿¡£Ä¿Ç°, ³£ÓõÄÁ×ËữÐÞÊε°°×·ÖÀ븻¼¯·½·¨Ö÷Òª°üÀ¨ÈýÀà: ÃâÒß³Áµí·¨¡¢Ç׺ÍÉ«Æ×·¨ºÍ»¯Ñ§ÑÜÉú·¨[50]¡£

¡¡¡¡½üÄêÀ´, µã»÷»¯Ñ§·´Ó¦ÔÚÁ×ËữÐÞÊε°°×µÄ¼ì²â·½ÃæÒ²·¢»ÓÁ˾޴ó×÷Óá£AkibaµÈ[51]·¢ÏÖÁËÒ»ÖÖ´øÕýµçºÉµÄË«ºËTb¢óÂçºÏÎïTb2-L, ¸ÃÂçºÏÎï½öÔÚÁ×ËáÀÒ°±Ëá´æÔÚÏÂÃ÷ÏÔ·¢¹â, ¿ÉÓÃÓÚÑ¡ÔñÐԵؼì²âÁ×ËáÀÒ°±Ëá(pTyr)¡£È»¶ø, µ±¶àëĵ×Îï´øÕýµçºÉʱ, Òò´æÔÚ¾²µç»¥³â×÷ÓÃ, ¼ì²âÁéÃô¶ÈÃ÷ÏÔ½µµÍ¡£Îª¿Ë·þ´Ëȱµã, ¸ÃÑо¿×éÌá³öÁËÒ»¸ö¸üÓŵķ½·¨, ÏòTb2-L·Ö×ÓÖÐÌí¼Ó´øÓÐȲ»ùµÄ»ùÍÅ, Éú³ÉÒ»ÖÖÐÂÐ͵ÄÂçºÏÎï̽ÕëTb2-Lc1yne; ͬʱ, ÏòÁ×Ëữ¶àëĵ×ÎïµÄ°ëë×°±Ëá²Ð»ùÉÏÒýÈëÒ»¸öµþµª»ùÍÅ, ×îºóÀûÓõþµª-Ȳ»ùµÄµã»÷»¯Ñ§·´Ó¦Ê¹ÂçºÏÎï̽ÕëÓëÁ×ËữµÄ¶àëĵ×ÎïʵÏÖ¹²¼Û½áºÏ¡£µ±¶àëĵ×Îï±»Á×Ëữʱ, »ùÓÚïçÏµÔªËØ·¢¹âµÄÐźŻᷢÉúÃ÷ÏԸıä, ´Ó¶øÊµÏÖø´ÙÀÒ°±ËáÁ×Ëữµ°°×µÄ¶¯Ì¬¼à²â¡£ÔÚÕâÏîÑо¿ÖÐ, ÀÒ°±ËáÁ×ËữëĶεÄÐźſÉͨ¹ýʱ¼ä·Ö±æ¹âÆ×ÇåÎúµØÓë±³¾°Çø·Ö¿ª, Ò×ÓÚ½øÐÐÏêϸµÄ¶¨Á¿·ÖÎö¡£×ÛÉÏ, ¸Ã·½·¨ÀûÓõã»÷»¯Ñ§·´Ó¦, Ö»Ð轫ÕâÖÖTb¢óÂçºÏÎïÌí¼Óµ½·´Ó¦»ìºÏÎïÖÐ, ¼´¿ÉʵÏÖ¶ÔÀÒ°±ËἤøµÄÁ×ËữºÍÈ¥Á×ËữµÄʵʱ¼à¿Ø, ÊÇÒ»ÖֲⶨDZÔÚµ°°×¼¤Ã¸ÒÖÖÆ¼ÁµÄÁ¼ºÃ¹¤¾ß, ¶ÔÒ©Î↑·¢µÄÑо¿¾ßÓÐÊ®·ÖÖØÒªµÄÍÆ½ø×÷Óá£

¡¡¡¡4¡¢ Õ¹ Íû

¡¡¡¡½üÄêÀ´, ½«µã»÷»¯Ñ§·´Ó¦ÓëSILAC¡¢BONCAT¡¢·Ö×ÓÓ¡¼£µÈ·½·¨Ïà½áºÏ, Ϊ¸ßͨÁ¿ÖÊÆ×·¨¼ì²âµ°°×ÖÊÌṩÁËÐÂ˼·¡£Í¨¹ýʹÓÿɷ¢Éúµã»÷·´Ó¦µÄ̽Õë, ¿É¶ÔÐÂÉú³Éµ°°×ºÍ·­ÒëºóÐÞÊε°°×½øÐпÉÊÓ»¯Ñо¿, ²¢ÊµÏÖµ°°×ÖÊ×éѧµÄ¶¨ÐÔÓ붨Á¿¼ì²â¡£¿¼Âǵ½µ°°×ÖÊÌÇ»ù»¯ÐÞÊκÍÁ×ËữÐÞÊεȷ­ÒëºóÐÞÊÎÔÚ¶àÖÖ¼²²¡ÖеĹؼü×÷ÓÃ, ¿ª·¢ÏàÓ¦µÄµ°°×ÌØÒìÐÔʶ±ð̽ÕëÖÁ¹ØÖØÒª¡£Ä¿Ç°, µã»÷»¯Ñ§·´Ó¦½öÔÚÉÙÊý¼¸ÖÖÀàÐ͵ĵ°°×ÖÊ·­ÒëºóÐÞÊÎÖеõ½Ó¦ÓÃ, ÀûÓõã»÷»¯Ñ§·´Ó¦¿ª·¢Õë¶ÔÆäËüÀàÐ͵ĵ°°×ÖÊ·­ÒëºóÐÞÊεÄÌØÒìÐÔ̽ÕëµÄÑо¿½«³ÉΪδÀ´Ò»¸öÖØÒªµÄÑо¿·½Ïò¡£Í¬Ê±, ¶ÔÓÚµ°°×Öʱê¼Ç¶øÑÔ, Ŀǰ½öÕë¶Ô¼×Áò°±ËῪ·¢ÁË¿ÉÖ±½Ó½øÐеã»÷·´Ó¦µÄС·Ö×ÓÀàËÆÎï, ¶øÕë¶ÔÆäËüÖÖÀàµÄ°±»ùËáɸѡ³öһϵÁÐÏàÓ¦ÀàËÆÎï, ½«¼«´óÍØÕ¹µã»÷»¯Ñ§·´Ó¦ÔÚµ°°×Öʶ¯Ì¬·ÖÎöÖеĹ㷺ӦÓᣴËÍâ, Í­´ß»¯µÄµã»÷»¯Ñ§·´Ó¦»á´øÀ´Ò»¶¨µÄÉúÎï¶¾ÐÔ, Òò´Ë, ¿ª·¢ÎÞ¶¾¸ßЧµÄÌåÄÚÐÂÐ͵ã»÷·´Ó¦ÈÔÈ»ÊÇδÀ´µã»÷»¯Ñ§·´Ó¦Ñо¿µÄÒ»¸öÈȵ㷽Ïò¡£

¡¡¡¡²Î¿¼ÎÄÏ×

¡¡¡¡[1] Williams K L,Gooley A A,Wilkins M R,Packer N H.J.Proteomics,2014,107:13-23
¡¡¡¡[2] Aslam B,Basit M,Nisar M A,Khurshid M,Rasool M H.J.Chromatogr.Sci.,2017,55(2):182-196
¡¡¡¡[3] Huang Z,Ma L G,Huang C H,Li Q F,Edouard C N.Proteomics,2017,17:6
¡¡¡¡[4] Shao S,Chen B,Cheng J,Wang C,Zhang Y,Shao L,Hu Y,Han Y,Han F,Li X.Biosens.Bioelectron.,2017,94:162-168
¡¡¡¡[5] Tse J,Wang Y,Zengeya T,Rozners E,Tan-Wilson A.Anal.Biochem.,2015,470:34-40
¡¡¡¡[6] Meyer J P,Adumeau P,Lewis J S,Zeglis B M.Bioconjugate Chem.,2016,27(12):2791-2807
¡¡¡¡[7] Güney A,Kasper L J,Georgi T,Sivakumar S,Jennifer R,Irina E,Iván C,Irena V,Erin S.RNA Biol.,2016,14(1):20-28
¡¡¡¡[8] Gao X,Hannoush R N.J.Am.Chem.Soc.,2014,136(12):4544-4550
¡¡¡¡[9] Suttapitugsakul S,Ulmer,L D,Jiang C,Sun F,Wu R.Anal.Chem.,2019,91(10):6934-6942
¡¡¡¡[10] Bertrand H C,Schaap M,Baird L,Georgakopoulos N D,Fowkes A,Thiollier C,Kachi H,Dinkova-Kostova A T,Wells G.J.Med.Chem.,2015,58(18):7186-7194
¡¡¡¡[11] Walczak S,Sikorski P J,Kasprzyk R,Kowalska J,Jemielity J.Org.Biomol.Chem.,2018,16(36):6741-6748
¡¡¡¡[12] Sherman E J,Lorenz D A,Garner A L.ACS Comb.Sci.,2019,21(7):522-527
¡¡¡¡[13] Eichelbaum K,Winter M,Berriel Diaz M,Herzig S,Krijgsveld J.Nat.Biotechnol.,2012,30(10):984-990
¡¡¡¡[14] Ma Y,Yates J R.Expert Rev.Proteomics,2018,15(7):545-554
¡¡¡¡[15] Wang J,Zhang J,Lee Y M.Nat.Protoc.,2017,12(2):279-288
¡¡¡¡[16] Teramoto H,Kojima K.Macromol.Biosci.,2015,15(6):719-727
¡¡¡¡[17] Dieterich D C,Lee J J,Link A J,Granmann J,Tirrell D A,Schuman E M.Nat.Protoc.,2007,2(3):532-540
¡¡¡¡[18] Serwa R A,Sekine E,Brown J,Teo S H C,Tate E W,O'Hare P.PLoS Pathog.,2019,15(10):e1007956
¡¡¡¡[19] Li Z,Zhu Y,Sun Y,Qin K,Liu W,Zhou W,Chen X.ACS Chem.Biol.,2016,11(12):3273-3277
¡¡¡¡[20] Franco M,D'Haeseleer P M,Branda S S,Liou M J,Haider Y,Segelke B W,El-Etr S H.Front.Cell Infect.Microbiol.,2018,8:370
¡¡¡¡[21] Dieterich D C,Link A J.Proc.Natl.Acad.Sci.USA,2006,103(25):9482-9487
¡¡¡¡[22] Shen W,Liu H H,Schiapparelli L,McClatchy D,He H,Yates J R,Cline H T.Cell Rep.,2014,6(4):737-747
¡¡¡¡[23] Howden A J,Geoghegan V,Katsch K,Efstathiou G,Bhushan B,Boutureira O,Thomas B,Trudgian D C,Kessler B M,Dieterich D C,Davis B G,Acuto O.Nat.Methods,2013,10(4):343-346
¡¡¡¡[24] Ma Y,McClatchy D B,Barkallah S,Wood W W,Yates J R.J.Proteome Res.,2017,16(6):2213-2220
¡¡¡¡[25] CUI Xiu-Yun,SUN Ning-Ning,XIE Xiao-Na,SUN Wan-Chun,ZHAO Qing,LIU Ning.Chinese J.Anal.Chem.,2018,46(11):1808-1813´ÞÐãÔÆ,ËïÄþÄþ,лÏþÄÈ,ËïÍò´º,ÕÔÇç,ÁõÄþ.·ÖÎö»¯Ñ§,2018,46(11):1808-1813
¡¡¡¡[26] Liu W,Brock A,Chen S,Chen S,Schultz P G.Nat.Methods,2007,4(3):239-244
¡¡¡¡[27] Wang L,Brock A,Herberich B,Schultz P G.Science,2001,292(5516):498-500
¡¡¡¡[28] Smits A H,Borrmann A,Roosjen Mark,Hest J C M,Vermeulen M.ACS Chem.Biol.,2016,11(2):3245-3250
¡¡¡¡[29] Woodley K T,Collins M O.Methods Mol.Biol.,2019,1977:71-82
¡¡¡¡[30] Brigidi G S,Bamji S X.J.Vis.Exp.,2013,(72):UNSP e50031
¡¡¡¡[31] Zhang X,Zhang Y,Fang C,Zhang L,Yang P,Wang C,Lu H.Anal.Chem.,2018,90:6161-6168
¡¡¡¡[32] FANG Cai-Yun,ZHANG Xiao-Qin,LU Hao-Jie.Chinese J.Anal.Chem.,2014,42(4):616-622·½²ÊÔÆ,ÕÅÏþÇÚ,½ºÀ½Ü.·ÖÎö»¯Ñ§,2014,42(4):616-622
¡¡¡¡[33] Thinon E,Fernandez J P,Molina H,Hang H C.J.Proteome Res.,2018,17(5):1907-1922
¡¡¡¡[34] Peng T,Hang H C.J.Am.Chem.Soc.,2015,137(2):556-559
¡¡¡¡[35] Yount J S,Moltedo B,Yang Y Y,Charron G,Moran T M,Lopez C B,Hang H C.Nat.Chem.Biol.,2010,6:610-614
¡¡¡¡[36] Teo C S H,Serwa R A,O'Hare P C.PLoS Pathog.,2016,12(10):e1005927
¡¡¡¡[37] Lai Z W,Nice E C,Oliver S L.Proteomics,2013,13(3-4):512-525
¡¡¡¡[38] Carsten G,Martin K,Yasmin G,Felix S,Andreas T,Lindhorst T K.Org.Biomol.Chem.,2013,11:4006
¡¡¡¡[39] Vajaria B N,Patel P S.Glycoconjugate J.,2017,34(2):147-156
¡¡¡¡[40] Wu Z L,Huang X,Burton A J,Swift K A.Carbohyd.Res.,2015,412:1-6
¡¡¡¡[41] Rong J,Han J,Dong L,Tan Y H,Yang H Q,Feng L S,Wang Q W,Meng R,Zhao J,Wang S Q.J.Am.Chem.Soc.,2014,136(50):17468-17476
¡¡¡¡[42] Herber J,Njavro J,Feederle R,Schepers U,Müller U C,Br?se S,Müller S A,Lichtenthaler S F.Mol.Cell.Proteomics,2018,17(8):1487-1501
¡¡¡¡[43] Song E,Mechref Y.Biomarkers Med.,2015,9(9):835-844
¡¡¡¡[44] Yang L,Nyalwidhe J O,Guo S,Drake R R,Semmes O J.Mol.Cell.Proteomics,2011,10(6):M110.007294
¡¡¡¡[45] Yang X H,Davision R T,Dong M.J.Am.Chem.Soc.,2018,140(33):10443-10446
¡¡¡¡[46] Vasapollo G,Sole R D,Mergola L,Lazzoi M R,Scardino A,Scorrano S,Mele G.Int.J.Mol.Sci.,2011,12(9):5908-5945
¡¡¡¡[47] Zhao T,Wang J,He J,Deng Q,Wang S.Biosens.Bioelectron.,2017,91:756-761
¡¡¡¡[48] Taracha A,Kotarba G,Wilanowski T.Postepy Biochem.,2017,63(2):137-142
¡¡¡¡[49] Singh V,Ram M,Kumar R,Prasad R,Roy B K,Singh K K.Protein J.,2017,36(1):1-6
¡¡¡¡[50] SHI Wen-Hao,TONG Meng-Sha,LI Kai,WANG Yu-Shen,DING Chen.Prog.Biochem.Biophys.,2018,45(12):1250-1258ʯÎĺÆ,ͯÃÎɯ,Àîâý,ÍõîÚ«|,¶¡è¡.ÉúÎﻯѧÓëÉúÎïÎïÀíÑо¿½øÕ¹,2018,45(12):1250-1258
¡¡¡¡[51] Akiba H,Sumaoka J,Tsumoto K,Komiyama M.Anal.Chem.,2015,87(7):3834-3840

×÷Õßµ¥Î»£º¼ªÁÖ´óѧµÚ¶þÒ½ÔºÖÐÐÄʵÑéÊÒ ¼ªÁÖ´óѧ¿ÚǻҽԺ ¼ªÁÖ´óѧÈËÊÞ¹²»¼²¡Ñо¿½ÌÓý²¿ÖصãʵÑéÊÒ
Ô­Îijö´¦£ºÕ¼·½Áá,¸ß˼Óî,лԪ¶°,ÕŽðÃú,ÀîÒã,ÁõÄþ.µã»÷»¯Ñ§·´Ó¦ÔÚµ°°×ÖÊ×éѧ·ÖÎöÖеÄÑо¿½øÕ¹[J].·ÖÎö»¯Ñ§,2020,48(04):431-438.
Ïà¹ØÄÚÈÝÍÆ¼ö
Ïà¹Ø±êÇ©£º
·µ»Ø£ºÉúÎﻯѧÂÛÎÄ