Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein

Manzi, Lucio and Barrow, Andrew and Hopper, Jonathan and Kaminska, Renata and Kleanthous, Colin and Robinson, Carol and Moses, John and Oldham, Neil (2017) Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein. Angewandte Chemie International Edition, 56 (47). pp. 14873-14877. ISSN 1521-3773

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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.

Item Type: Article
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.
Keywords: carbene footprinting; integral membrane proteins; mass spectrometry; protein labelling; protein structures
Schools/Departments: University of Nottingham, UK > Faculty of Science > School of Chemistry
Identification Number: https://doi.org/10.1002/anie.201708254
Depositing User: Oldham, Dr Neil J
Date Deposited: 03 Oct 2017 11:45
Last Modified: 27 Sep 2018 04:30
URI: http://eprints.nottingham.ac.uk/id/eprint/46932

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