Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein
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-b...
| Main Authors: | , , , , , , , |
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| Format: | Article |
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Wiley-VCH Verlag
2017
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| Online Access: | https://eprints.nottingham.ac.uk/46932/ |
| _version_ | 1848797430896656384 |
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| author | Manzi, Lucio Barrow, Andrew Hopper, Jonathan Kaminska, Renata Kleanthous, Colin Robinson, Carol Moses, John Oldham, Neil |
| author_facet | Manzi, Lucio Barrow, Andrew Hopper, Jonathan Kaminska, Renata Kleanthous, Colin Robinson, Carol Moses, John Oldham, Neil |
| author_sort | Manzi, Lucio |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-14T20:03:46Z |
| format | Article |
| id | nottingham-46932 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:03:46Z |
| publishDate | 2017 |
| publisher | Wiley-VCH Verlag |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-469322020-05-04T19:18:55Z https://eprints.nottingham.ac.uk/46932/ 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 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. Wiley-VCH Verlag 2017-11-20 Article PeerReviewed 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 carbene footprinting; integral membrane proteins; mass spectrometry; protein labelling; protein structures http://onlinelibrary.wiley.com/doi/10.1002/anie.201708254/abstract?campaign=wolacceptedarticle doi:10.1002/anie.201708254 doi:10.1002/anie.201708254 |
| spellingShingle | carbene footprinting; integral membrane proteins; mass spectrometry; protein labelling; protein structures Manzi, Lucio Barrow, Andrew Hopper, Jonathan Kaminska, Renata Kleanthous, Colin Robinson, Carol Moses, John Oldham, Neil Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| title | Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| title_full | Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| title_fullStr | Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| title_full_unstemmed | Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| title_short | Carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| title_sort | carbene footprinting reveals binding interfaces of a multimeric membrane spanning protein |
| topic | carbene footprinting; integral membrane proteins; mass spectrometry; protein labelling; protein structures |
| url | https://eprints.nottingham.ac.uk/46932/ https://eprints.nottingham.ac.uk/46932/ https://eprints.nottingham.ac.uk/46932/ |