首页|Encouraging Solution to the Problem of Synthesizing Protein α-Thioester

Encouraging Solution to the Problem of Synthesizing Protein α-Thioester

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Expressed protein ligation(EPL)provides a powerful tool to access large-size proteins with precise structures.Existing methods for constructing the critical protein thioester for EPL have predominantly relied on the recombinant intein fusion expressed in Escherichia coli(E.coli).Despite its powerful applications,the expression of thioester derived from eukaryotic protein in E.coli inherently suffers from its limited solubility,the inactivity of intein,premature hydrolysis and low yields.To overcome these obstacles,we present herein the facile one-flask synthesis of inaccessible protein α-thioester via a SUMO-protein-intein(SPI)sandwich model.The utility of SUMO enhances the protein fusion yield and solubility,prevents premature hydrolysis and simplifies the purification process.The inaccessi-ble protein thioester with internal Cys residues can be readily produced and is compatible with the EPL-desulfurization protocol used to prepare complex proteins,which is otherwise difficult to obtain using traditional methods.Its utility has been highlighted through the synthesis of human granulocyte colony-stimulating factor(G-CSF).

DesulfurizationGlycoproteinsGlycopeptidesProtein expressionPeptide thioester

Xinliang Liu、Zijun Gao、Jie Zhao、Farong Ye、Ping Huang、Ping Wang

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Center for Chemical Glycobiology,Shanghai Key Laboratory for Molecular Engineering of Chiral Drugs,Frontiers Science Center for Transformative Molecules,School of Chemistry and Chemical Engineering,Zhangjiang Institute for Advanced Study,Shanghai Jiao Tong University,Shanghai 200240,China

Shenzhen Research Institute of Shanghai Jiao Tong University,Shenzhen,Guangdong 518057,China

National Science Fund for Distinguished Young ScholarsInterdisciplinary Program of Shanghai Jiao Tong UniversityNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaNational Natural Science Foundation of ChinaShanghai Pilot Program for Basic Research Shanghai Jiao Tong UniversitySpecial Projects of the Central Government in Guidance of Local Science and Technology DevelopmentChina Postdoctoral Science Foundation

222257012023010222077080922533022227512221TQ14002102021Szvup0772023M732214

2024

10.1002/cjoc.202300762

中国化学(英文版)
中国化学会 上海有机化学研究所

中国化学(英文版)

CSTPCD
影响因子：0.848
ISSN：1001-604X
年,卷(期)：2024.42(10)