Biosynthesis of peptide fragments of eukaryotic GPCRs in Escherichia coli by directing expression into inclusion bodies

被引：8

作者：

Cohen, Leah S. ^{[1
,2
]}

Becker, Jeffrey M. ^{[3
]}

Naider, Fred ^{[1
,2
]}

机构：

[1] CUNY Coll Staten Isl, Dept Chem, Staten Isl, NY 10314 USA

[2] CUNY, Grad Ctr, Dept Biochem, New York, NY 10016 USA

[3] Univ Tennessee, Dept Microbiol, Knoxville, TN 37996 USA

来源：

JOURNAL OF PEPTIDE SCIENCE | 2010年 / 16卷 / 05期

关键词：

biosynthesis; G protein-coupled receptor; GPCR; peptide fragments; isotopic labeling; membrane peptides; PROTEIN-COUPLED RECEPTOR; TRANSMEMBRANE DOMAINS; NMR CHARACTERIZATION; BIOPHYSICAL ANALYSIS; PURIFICATION; HIV-1; CB2; SPECTROSCOPY; VPU;

D O I：

10.1002/psc.1222

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Biosynthesis of peptides in heterologous systems is often a prerequisite to biophysical analyses. Large amounts of peptides, in particular portions of membrane proteins, are needed to optimize conditions for secondary and tertiary structure analysis. Chemical synthesis of these peptides is limited by their high hydrophobicity and also due to the need to incorporate isotopic labels for high resolution NMR analysis. In this protocol, we describe a method for the heterologous expression and purification of unlabeled and isotopically labeled peptide fragments of Ste2p, an integral membrane G protein-coupled receptor. Copyright (C) 2010 European Peptide Society and John Wiley & Sons, Ltd.

引用

页码：213 / 218

页数：6

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