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Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein

Manzi, Lucio; Barrow, Andrew; Hopper, Jonathan; Kaminska, Renata; Kleanthous, Colin; Robinson, Carol; Moses, John; Oldham, Neil

Authors

Lucio Manzi

Andrew Barrow

Jonathan Hopper

Renata Kaminska

Colin Kleanthous

Carol Robinson

John Moses

Neil Oldham



Abstract

Mapping the interaction sites between membrane spanning proteins is a key challenge in structural biology. In this study a carbene footprinting approach is developed and applied to identify the interfacial sites of a trimeric, integral membrane protein, OmpF, solubilised in micelles. The diazirine-based footprinting probe is effectively sequestered by, and incorporated into, the micelles leading to efficient labelling of the membrane-spanning regions of the protein upon irradiation at 349 nm. Areas associated with protein-protein interactions between the trimer subunits remained unlabelled, thus revealing their location.

Journal Article Type Article
Publication Date Nov 20, 2017
Journal Angewandte Chemie International Edition
Print ISSN 1433-7851
Electronic ISSN 1521-3773
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 56
Issue 47
APA6 Citation Manzi, L., Barrow, A., Hopper, J., Kaminska, R., Kleanthous, C., Robinson, C., …Oldham, N. (2017). Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein. Angewandte Chemie International Edition, 56(47), doi:10.1002/anie.201708254
DOI https://doi.org/10.1002/anie.201708254
Keywords carbene footprinting; integral membrane proteins; mass spectrometry; protein labelling; protein structures
Publisher URL http://onlinelibrary.wiley.com/doi/10.1002/anie.201708254/abstract?campaign=wolacceptedarticle
Copyright Statement Copyright information regarding this work can be found at the following address: http://eprints.nottingh.../end_user_agreement.pdf
Additional Information This is the peer reviewed version of the following article: Manzi, L., Barrow, A., Hopper, J., Kaminska, R., Kleanthous, C., Robinson, C., Moses, J. and Oldham, N. J. (), Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein. Angew. Chem. Int. Ed., which has been published in final form at http://dx.doi.org/10.1002/anie.201708254. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.

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OmpF Footprinting manuscript.pdf (1.2 Mb)
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Copyright Statement
Copyright information regarding this work can be found at the following address: http://eprints.nottingham.ac.uk/end_user_agreement.pdf





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