Ion mobility-mass spectrometry reveals conformational flexibility in the deubiquitinating enzyme USP5

Scott, Daniel and 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

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Abstract

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.

Item Type: Article
Keywords: electrospray ionisation, ion mobility-mass spectrometry, protein conformation, ubiquitin specific protease 5
Schools/Departments: University of Nottingham UK Campus > Faculty of Medicine and Health Sciences > School of Life Sciences > School of Biomedical Sciences
University of Nottingham UK Campus > Faculty of Science > School of Chemistry
Identification Number: https://doi.org/10.1002/pmic.201400457
Related URLs:
URLURL Type
http://www.interscience.wiley.com/Publisher
Depositing User: Oldham, Dr Neil J
Date Deposited: 01 Jul 2016 07:21
Last Modified: 14 Sep 2016 09:06
URI: http://eprints.nottingham.ac.uk/id/eprint/34559

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