Site-specific protein modification by genetic encoded disulfide compatible thiols  

作　　者：Xinyu Ling Heqi Chen Wei Zheng Liying Chang Yong Wang Tao Liu 

机构地区：[1]State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,Peking University

出　　处：《Chinese Chemical Letters》2020年第1期163-166,共4页中国化学快报（英文版）

基　　金：financially supported by National Key Research and Development Program of China (No.2016YFA0201400);the National Natural Science Foundation of China (No.21778005);Peking University Health Science Center (Nos.BMU20160537 andBMU2017QQ006);the Youth Thousand-Talents Program of China for support

摘　　要：Cysteine chemistry provides a low cost and convenient way for site-specific protein modification.However,recombinant expression of disulfide bonding containing protein with unpaired cysteine is technically challenging and the resulting protein often suffers from significantly reduced yield and activity.Here we used genetic code expansion technique to introduce a surface exposed self-paired dithiol functional group into proteins,which can be selectively reduced to afford active thiols.Two compounds containing self-paired disulfides were synthesized,and their genetic incorporations were validated using green fluorescent proteins(GFP).The compatibility of these self-paired di-thiols with natural disulfide bond was demonstrated using antibody fragment to afford site-specifically labeled antibody.This work provides another valuable building block into the chemical tool-box for site-specific labeling of proteins containing internal disulfides.Cysteine chemistry provides a low cost and convenient way for site-specific protein modification.However,recombinant expression of disulfide bonding containing protein with unpaired cysteine is technically challenging and the resulting protein often suffers from significantly reduced yield and activity.Here we used genetic code expansion technique to introduce a surface exposed self-paired dithiol functional group into proteins,which can be selectively reduced to afford active thiols.Two compounds containing self-paired disulfides were synthesized,and their genetic incorporations were validated using green fluorescent proteins(GFP).The compatibility of these self-paired di-thiols with natural disulfide bond was demonstrated using antibody fragment to afford site-specifically labeled antibody.This work provides another valuable building block into the chemical tool-box for site-specific labeling of proteins containing internal disulfides.

关 键 词：Cysteine chemistry Genetic code expansion Protein modification Biorthogonal chemistry Disulfide bond 

分 类 号：TQ931[化学工程]