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: High Magnetic Field Laboratory,Chinese Academy of Sciences, University of Science and Technology of
• 出 处: Science China(Chemistry)
• 发表日期: 2013年09期
• 关键词: ligation conjugate Fmoc equiv assembled sequential expensive reagent ubiquitin c

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 throug

The C-terminal conjugate of ubiquitin with 7-amino-4-methylcoumarin (Ub-AMC) is an important probe for fluorescence based evaluation 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 the expected biological functions throug