Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5
Many proteins exhibit conformation flexibility as part of their biological function, whether through the presence of a series of well-defined states or by the existence of intrinsic disorder. Ion mobility spectrometry, in combination with MS (IM–MS), offers a rapid and sensitive means of probing ens...
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| Format: | Article |
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Wiley
2015
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| Online Access: | https://eprints.nottingham.ac.uk/34559/ |
| _version_ | 1848794882311716864 |
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| author | Scott, Daniel Layfield, Robert Oldham, Neil J. |
| author_facet | Scott, Daniel Layfield, Robert Oldham, Neil J. |
| author_sort | Scott, Daniel |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Many proteins exhibit conformation flexibility as part of their biological function, whether through the presence of a series of well-defined states or by the existence of intrinsic disorder. Ion mobility spectrometry, in combination with MS (IM–MS), offers a rapid and sensitive means of probing ensembles of protein structures through measurement of gas-phase collisional cross sections. We have applied IM–MS analysis to the multidomain deubiquitinating enzyme ubiquitin specific protease 5 (USP5), which is believed to exhibit significant conformational flexibility. Native ESI–MS measurement of the 94-kDa USP5 revealed two distinct charge-state distributions: [M + 17H]+ to [M + 21H]+ and [M + 24H]+ to [M + 29H]+. The collisional cross sections of these ions revealed clear groupings of 52 ± 4 nm2 for the lower charges and 66 ± 6 nm2 for the higher charges. Molecular dynamics simulation of a compact form of USP5, based on a crystal structure, produced structures of 53–54 nm2 following 2 ns in the gas phase, while simulation of an extended form (based on small-angle X-ray scattering data) led to structures of 64 nm2. These data demonstrate that IM–MS is a valuable tool in studying proteins with different discrete conformational states. |
| first_indexed | 2025-11-14T19:23:15Z |
| format | Article |
| id | nottingham-34559 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T19:23:15Z |
| publishDate | 2015 |
| publisher | Wiley |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-345592020-05-04T17:04:52Z https://eprints.nottingham.ac.uk/34559/ Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 Scott, Daniel Layfield, Robert Oldham, Neil J. Many proteins exhibit conformation flexibility as part of their biological function, whether through the presence of a series of well-defined states or by the existence of intrinsic disorder. Ion mobility spectrometry, in combination with MS (IM–MS), offers a rapid and sensitive means of probing ensembles of protein structures through measurement of gas-phase collisional cross sections. We have applied IM–MS analysis to the multidomain deubiquitinating enzyme ubiquitin specific protease 5 (USP5), which is believed to exhibit significant conformational flexibility. Native ESI–MS measurement of the 94-kDa USP5 revealed two distinct charge-state distributions: [M + 17H]+ to [M + 21H]+ and [M + 24H]+ to [M + 29H]+. The collisional cross sections of these ions revealed clear groupings of 52 ± 4 nm2 for the lower charges and 66 ± 6 nm2 for the higher charges. Molecular dynamics simulation of a compact form of USP5, based on a crystal structure, produced structures of 53–54 nm2 following 2 ns in the gas phase, while simulation of an extended form (based on small-angle X-ray scattering data) led to structures of 64 nm2. These data demonstrate that IM–MS is a valuable tool in studying proteins with different discrete conformational states. Wiley 2015-03-09 Article PeerReviewed Scott, Daniel, Layfield, Robert and Oldham, Neil J. (2015) Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5. Proteomics, 15 (16). pp. 2835-2841. ISSN 1615-9861 electrospray ionisation ion mobility-mass spectrometry protein conformation ubiquitin specific protease 5 http://onlinelibrary.wiley.com/doi/10.1002/pmic.201400457/abstract doi:10.1002/pmic.201400457 doi:10.1002/pmic.201400457 |
| spellingShingle | electrospray ionisation ion mobility-mass spectrometry protein conformation ubiquitin specific protease 5 Scott, Daniel Layfield, Robert Oldham, Neil J. Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 |
| title | Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 |
| title_full | Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 |
| title_fullStr | Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 |
| title_full_unstemmed | Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 |
| title_short | Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5 |
| title_sort | ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme usp5 |
| topic | electrospray ionisation ion mobility-mass spectrometry protein conformation ubiquitin specific protease 5 |
| url | https://eprints.nottingham.ac.uk/34559/ https://eprints.nottingham.ac.uk/34559/ https://eprints.nottingham.ac.uk/34559/ |