1. Academic Validation
  2. In situ EPR spectroscopy of a bacterial membrane transporter using an expanded genetic code

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

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].
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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