Dissemin is shutting down on January 1st, 2025

Published in

Wiley, Protein Science, 5(5), p. 981-981, 1996

DOI: 10.1002/pro.5560050522

Wiley, Protein Science, 1(5), p. 174-177, 1996

DOI: 10.1002/pro.5560050123

Links

Tools

Export citation

Search in Google Scholar

Rapid screening for structural integrity of expressed proteins by heteronuclear NMR spectroscopy.

Journal article published in 1996 by Angela M. Gronenborn ORCID, G. Marius Clore, G. Marius Clore
This paper is made freely available by the publisher.
This paper is made freely available by the publisher.

Full text: Download

Green circle
Preprint: archiving allowed
Upload
Orange circle
Postprint: archiving restricted
Upload
Red circle
Published version: archiving forbidden
Policy details
Data provided by SHERPA/RoMEO

Abstract

A simple and rapid method based on 15N labeling and 1H-15N heteronuclear single quantum coherence spectroscopy is presented to directly assess the structural integrity of overexpressed proteins in crude Escherichia coli extracts without the need for any purification. The method is demonstrated using two different expression systems and two different proteins, the B1 immunoglobulin-binding domain of streptococcal protein G (56 residues) and human interleukin-1 beta (153 residues). It is shown that high quality 1H-15N correlation spectra, recorded in as little as 15 min and displaying only cross-peaks arising from the overexpressed protein of interest, can be obtained from crude E. coli extracts.