Manzi, L., Barrow, A. S., Scott, D., Layfield, R., Wright, T. G., Moses, J. E., Oldham, N. J. (November 2016) Carbene footprinting accurately maps binding sites in protein-ligand and protein-protein interactions. Nat Commun, 7. p. 13288. ISSN 2041-1723
Abstract
Specific interactions between proteins and their binding partners are fundamental to life processes. The ability to detect protein complexes, and map their sites of binding, is crucial to understanding basic biology at the molecular level. Methods that employ sensitive analytical techniques such as mass spectrometry have the potential to provide valuable insights with very little material and on short time scales. Here we present a differential protein footprinting technique employing an efficient photo-activated probe for use with mass spectrometry. Using this methodology the location of a carbohydrate substrate was accurately mapped to the binding cleft of lysozyme, and in a more complex example, the interactions between a 100�kDa, multi-domain deubiquitinating enzyme, USP5 and a diubiquitin substrate were located to different functional domains. The much improved properties of this probe make carbene footprinting a viable method for rapid and accurate identification of protein binding sites utilizing benign, near-UV photoactivation.
| Item Type: | Paper |
|---|---|
| Additional Information: | 2041-1723 Manzi, Lucio Barrow, Andrew S Scott, Daniel Layfield, Robert Wright, Timothy G Moses, John E Oldham, Neil J BB/F019297/1/Biotechnology and Biological Sciences Research Council/United Kingdom Journal Article Research Support, Non-U.S. Gov't Nat Commun. 2016 Nov 16;7:13288. doi: 10.1038/ncomms13288. |
| CSHL Authors: | |
| Communities: | CSHL labs > Moses lab |
| Depositing User: | Matthew Dunn |
| Date: | 16 November 2016 |
| Date Deposited: | 08 Jan 2021 18:06 |
| Last Modified: | 08 Jan 2021 18:06 |
| PMCID: | PMC5116083 |
| URI: | https://repository.cshl.edu/id/eprint/39559 |
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