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Biosynthesis of peptide fragments of eukaryotic GPCRs in Escherichia coli by directing expression into inclusion bodies
Author(s) -
Cohen Leah S.,
Becker Jeffrey M.,
Naider Fred
Publication year - 2010
Publication title -
journal of peptide science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.475
H-Index - 66
eISSN - 1099-1387
pISSN - 1075-2617
DOI - 10.1002/psc.1222
Subject(s) - g protein coupled receptor , heterologous expression , heterologous , inclusion bodies , biosynthesis , biochemistry , escherichia coli , chemistry , peptide , membrane protein , peptide synthesis , integral membrane protein , membrane , computational biology , receptor , recombinant dna , biology , gene
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 © 2010 European Peptide Society and John Wiley & Sons, Ltd.