Chemical synthesis of Ub-AMC via ligation of peptide hydrazides  

作　　者：LIANG Jun FANG GeMin HUANG XiuLiang MEI ZiQing LI Juan TIAN ChangLin LIU Lei 

机构地区：[1]High Magnetic Field Laboratory,Chinese Academy of Sciences [2]University of Science and Technology of China [3]Tsinghua-Peking Center for Life Sciences,Department of Chemistry,Tsinghua University [4]Ministry of Education Protein Science Laboratory,Center for Structural Biology,School of Life Sciences,Tsinghua University

出　　处：《Science China Chemistry》2013年第9期1301-1306,共6页中国科学（化学英文版）

基　　金：National Basic Research Program of China (973 program, 2013CB932800);the National Natural Science Foundation of China (NSFC, 31100524 to M.Z., 31170817 for C.T., and 20972148 to L.L.)

摘　　要：The C-terminal conjugate of ubiquitin with 7-amino-4-methylcoumarin （Ub-AMC） is an important probe for fluorescencebased analysis of deubiquitinating enzyme （DUB） activity. It is important to develop more efficient methods for the preparation of Ub-AMC because the currently available technology is still expensive for scaled-up production. In the present work we report an efficient strategy for total chemical synthesis of Ub-AMC through ligation of peptide hydrazides. Three peptide segments are assembled via N-to-C sequential ligation and the resulting product is converted to Ub-AMC via TCEP-mediated desulfurization. The synthetic Ub-AMC is shown to have expected biological functions througThe C-terminal conjugate of ubiquitin with 7-amino-4-methylcoumarin (Ub-AMC) is an important probe for fluorescencebased analysis of deubiquitinating enzyme (DUB) activity. It is important to develop more efficient methods for the preparation of Ub-AMC because the currently available technology is still expensive for scaled-up production. In the present work we report an efficient strategy for total chemical synthesis of Ub-AMC through ligation of peptide hydrazides. Three peptide segments are assembled via N-to-C sequential ligation and the resulting product is converted to Ub-AMC via TCEP-mediated desulfurization. The synthetic Ub-AMC is shown to have expected biological functions

关 键 词：h the measurement of its DUB activity.ligation of peptide hydrazides peptide segment condensation deubiquitinating enzymes UBIQUITIN 

分 类 号：O621.3[理学—有机化学]