Chemistry and Physics Faculty Articles

Title

Distance Measurement on an Endogenous Membrane Transporter in E. coli Cells and Native Membranes Using EPR Spectroscopy

Document Type

Article

Publication Date

5-18-2015

Publication Title

Angewandte Chemie International Edition

Keywords

In-cell spectroscopy, Membrane proteins, EPR spectroscopy, Spin labeling, Vitamin B12 transporter

ISSN

1433-7851

Volume

54

Issue/No.

21

First Page

6196

Last Page

6199

Abstract

Membrane proteins may be influenced by the environment, and they may be unstable in detergents or fail to crystallize. As a result, approaches to characterize structures in a native environment are highly desirable. Here, we report a novel general strategy for precise distance measurements on outer membrane proteins in whole Escherichia coli cells and isolated outer membranes. The cobalamin transporter BtuB was overexpressed and spin‐labeled in whole cells and outer membranes and interspin distances were measured to a spin‐labeled cobalamin using pulse EPR spectroscopy. A comparative analysis of the data reveals a similar interspin distance between whole cells, outer membranes, and synthetic vesicles. This approach provides an elegant way to study conformational changes or protein–protein/ligand interactions at surface‐exposed sites of membrane protein complexes in whole cells and native membranes, and provides a method to validate outer membrane protein structures in their native environment.

Comments

©2015 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim

Additional Comments

Deutsche Forschungsgemeinschaft grant #: SFB 807; NIH grant #: GM035215

DOI

10.1002/anie.201501086

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Peer Reviewed

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