In situ EPR spectroscopy of a bacterial membrane transporter using an expanded genetic code

Anandi Kugele; Sophie Ketter; Bjarne Silkenath; Valentin Wittmann; Benesh Joseph; Malte Drescher

doi:10.1039/D1CC04612H

In situ EPR spectroscopy of a bacterial membrane transporter using an expanded genetic code†

Anandi Kugele,

^a Sophie Ketter,^b Bjarne Silkenath,

^a Valentin Wittmann,

^a Benesh Joseph

*^b and Malte Drescher

*^a

Author affiliations

* Corresponding authors

^a Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz, Universitätsstraße 10, 78457 Konstanz, Germany
E-mail: malte.drescher@uni-konstanz.de

^b Institute of Biophysics, Department of Physics & The Center for Biomolecular Magnetic Resonance (BMRZ), Goethe University Frankfurt, Max-von-Laue-Str. 1, 60438 Frankfurt/Main, Germany
E-mail: joseph@biophysik.uni-frankfurt.de

Abstract

The membrane transporter BtuB is site-directedly spin labelled on the surface of living Escherichia coli via Diels–Alder click chemistry of the genetically encoded amino acid SCO-L-lysine. The previously introduced photoactivatable nitroxide PaNDA prevents off-target labelling, is used for distance measurements, and the temporally shifted activation of the nitroxide allows for advanced experimental setups. This study describes significant evolution of Diels–Alder-mediated spin labelling on cellular surfaces and opens up new vistas for the the study of membrane proteins.

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DOI: https://doi.org/10.1039/D1CC04612H
Article type: Communication
Submitted: 19 Aug 2021
Accepted: 03 Nov 2021
First published: 18 Nov 2021
This article is Open Access

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Chem. Commun., 2021,57, 12980-12983

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In situ EPR spectroscopy of a bacterial membrane transporter using an expanded genetic code

A. Kugele, S. Ketter, B. Silkenath, V. Wittmann, B. Joseph and M. Drescher, Chem. Commun., 2021, 57, 12980 DOI: 10.1039/D1CC04612H

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

Chemical Communications

In situ EPR spectroscopy of a bacterial membrane transporter using an expanded genetic code†

Abstract

Supplementary files

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In situ EPR spectroscopy of a bacterial membrane transporter using an expanded genetic code

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