Carbene Footprinting Reveals Binding Interfaces of a Multimeric Membrane‐Spanning Protein. (20th October 2017)

• Record Type: Journal Article
• Authors: Manzi, Lucio; Barrow, Andrew S.; Hopper, Jonathan T. S.; Kaminska, Renata; Kleanthous, Colin; Robinson, Carol V.; Moses, John E.; Oldham, Neil J.
• Is Part Of: Angewandte Chemie. Volume 129:Number 47(2017)
• Page Start: 15069
• Page End: 15073
• Publication Date: 2017-10-20
• Subjects: Carben-Footprinting -- Integrale Membranproteine -- Massenspektrometrie -- Proteinmarkierung -- Proteinstrukturen; Chemistry -- Periodicals; 540
• DOI: 10.1002/ange.201708254
• Languages: English
• ISSNs: 0044-8249

Abstract

Abstract: Mapping the interaction sites between membrane‐spanning proteins is a key challenge in structural biology. In this study a carbene‐footprinting approach was 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, thus 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.