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

  • Chem Commun (Camb). 2021 Dec 3;57(96):12980-12983. doi: 10.1039/d1cc04612h.
Anandi Kugele  1 ,  Sophie Ketter  2 ,  Bjarne Silkenath  1 ,  Valentin Wittmann  1 ,  Benesh Joseph  2 ,  Malte Drescher  1
Affiliations
  • 1. Department of Chemistry and Konstanz Research School Chemical Biology (KoRS-CB), University of Konstanz, Universitätsstraße 10, 78457 Konstanz, Germany. [email protected].
  • 2. 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. [email protected].
PMID: 34792069 DOI: 10.1039/d1cc04612h
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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