Efficient cellular solid-state NMR of membrane proteins by targeted protein labeling

Lindsay A. Baker; Mark Daniëls; Elwin A W Van Der Cruijsen; Gert E. Folkers; Marc Baldus

doi:10.1007/s10858-015-9936-5

Efficient cellular solid-state NMR of membrane proteins by targeted protein labeling

Lindsay A. Baker, Mark Daniëls, Elwin A W Van Der Cruijsen, Gert E. Folkers, Marc Baldus^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

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 language	English
Pages (from-to)	199-208
Number of pages	10
Journal	Journal of Biomolecular NMR
Volume	62
Issue number	2
DOIs	https://doi.org/10.1007/s10858-015-9936-5
Publication status	Published - 4 Jun 2015

Keywords

Cellular membranes
Membrane protein
Protein expression
Solid-state NMR

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abstract = "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.",

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AU - Daniëls, Mark

AU - Van Der Cruijsen, Elwin A W

AU - Folkers, Gert E.

AU - Baldus, Marc

PY - 2015/6/4

Y1 - 2015/6/4

N2 - 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.

AB - 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.

KW - Cellular membranes

KW - Membrane protein

KW - Protein expression

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Efficient cellular solid-state NMR of membrane proteins by targeted protein labeling

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Keywords

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