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Solid-state NMR spectroscopy (ssNMR) has made significant progress towards the study of membrane proteins in their native cellular membranes. However, reduced spectroscopic sensitivity and high background signal levels can complicate these experiments. Here, we describe a method for ssNMR to specifically label a single protein by repressing endogenous protein expression with rifampicin. Our results demonstrate that treatment of E. coli with rifampicin during induction of recombinant membrane protein expression reduces background signals for different expression levels and improves sensitivity in cellular membrane samples. Further, the method reduces the amount of time and resources needed to produce membrane protein samples, enabling new strategies for studying challenging membrane proteins by ssNMR.

Original publication

DOI

10.1007/s10858-015-9936-5

Type

Journal article

Journal

Journal of biomolecular NMR

Publication Date

06/2015

Volume

62

Pages

199 - 208

Addresses

NMR Spectroscopy, Department of Chemistry, Faculty of Science, Bijvoet Center for Biomolecular Research, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands.

Keywords

Streptomyces lividans, Escherichia coli, Rifampin, Proteolipids, Bacterial Proteins, Escherichia coli Proteins, Membrane Transport Proteins, Potassium Channels, Recombinant Proteins, Nuclear Magnetic Resonance, Biomolecular, Sensitivity and Specificity, Cloning, Molecular, Protein Conformation