Chemical synthesis of Ub-AMC via ligation of peptide hydrazides

• Published in: Science China. Chemistry Vol. 56; no. 9; pp. 1301 - 1306
• Main Authors: Liang, Jun, Fang, GeMin, Huang, XiuLiang, Mei, ZiQing, Li, Juan, Tian, ChangLin, Liu, Lei
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.09.2013; Springer Nature B.V
• Subjects: Chemical synthesis; Chemistry; Chemistry and Materials Science; Chemistry/Food Science; Hydrazides; 化学合成; 合成肽; 泛素化; 活性分析; 生物学功能; 甲基香豆素; 结扎; 酰肼; ligation of peptide hydrazides; peptide segment condensation; ubiquitin; deubiquitinating enzymes
• ISSN: 1674-7291; 1869-1870
• DOI: 10.1007/s11426-013-4885-x

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