Carbene footprinting reveals binding interfaces of a multimeric membrane spanning proteinTools 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 Full text not available from this repository.
Official URL: http://onlinelibrary.wiley.com/doi/10.1002/anie.201708254/abstract?campaign=wolacceptedarticle
AbstractMapping 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.
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