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

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...

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Detalles Bibliográficos
Autores principales: Kugele, Anandi, Ketter, Sophie, Silkenath, Bjarne, Wittmann, Valentin, Joseph, Benesh, Drescher, Malte
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Royal Society of Chemistry 2021
Materias:
Chemistry
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8640571/
https://www.ncbi.nlm.nih.gov/pubmed/34792069
http://dx.doi.org/10.1039/d1cc04612h
Descripción
Sumario: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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