Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein

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Manzi, Lucio, Barrow, Andrew, Hopper, Jonathan, Kaminska, Renata, Kleanthous, Colin, Robinson, Carol, 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
RIS ID: https://nottingham-repository.worktribe.com/output/895876
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: 10.1002/anie.201708254
Depositing User: Oldham, Dr Neil J
Date Deposited: 03 Oct 2017 11:45
Last Modified: 04 May 2020 19:18
URI: https://eprints.nottingham.ac.uk/id/eprint/46932

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