使用内含肽重折叠策略表达N端半胱氨酸蛋白片段

Expression of N-terminal Cys-protein fragments using an intein refolding strategy.

作者信息

Hackenberger Christian P R, Chen Mark M, Imperiali Barbara

机构信息

Massachusetts Institute of Technology, Department of Chemistry and Department of Biology, 77 Massachusetts Ave. 18-590, Cambridge, MA 02139, USA.

出版信息

Bioorg Med Chem. 2006 Jul 15;14(14):5043-8. doi: 10.1016/j.bmc.2006.03.003. Epub 2006 Mar 22.

DOI:10.1016/j.bmc.2006.03.003

PMID:16554163

Abstract

The inclusion body expression and refolding of a pH-sensitive intein fusion protein (Ssp DnaB intein) delivered sufficient quantities of an N-terminal Cys-polypeptide for native chemical ligations. This strategy circumvents premature intein cleavage under expression conditions and allows the expression and purification of proteins with uncertain solubility properties. The expressed protein resembles the C-terminal portion of the amphiphilic immunity protein Im7, which can be ligated to synthetic thioesters to yield synthetic protein analogues for protein folding studies.

摘要

一种pH敏感型内含肽融合蛋白（Ssp DnaB内含肽）的包涵体表达及重折叠，为天然化学连接提供了足够量的N端半胱氨酸多肽。该策略避免了在表达条件下内含肽的过早切割，并允许表达和纯化具有不确定溶解特性的蛋白质。所表达的蛋白质类似于两亲性免疫蛋白Im7的C端部分，其可与合成硫酯连接，以产生用于蛋白质折叠研究的合成蛋白质类似物。