Exploring the conformational landscape and stability of Aurora A using ion-mobility mass spectrometry and molecular modelling

Tomlinson, Lauren J, Batchelor, Matthew, Sarsby, Joscelyn, Byrne, Dominic P, Brownridge, Philip, Bayliss, Richard, Eyers, Patrick A ORCID: 0000-0002-9220-2966 and Eyers, Claire E ORCID: 0000-0002-3223-5926 (2022) Exploring the conformational landscape and stability of Aurora A using ion-mobility mass spectrometry and molecular modelling. Journal of the American Society for Mass Spectrometry, 33 (3). pp. 420-435.

Access the full-text of this item by clicking on the Open Access link.

Official URL: http://dx.doi.org/10.1021/jasms.1c00271

Abstract

Protein kinase inhibitors are highly effective in treating diseases driven by aberrant kinase signalling, and as chemical tools to help dissect the cellular roles of kinase signalling complexes. Evaluating the effects of binding of small molecule inhibitors on kinase conformational dynamics can assist in understanding both inhibition and resistance mechanisms. Using gas-phase ion mobility-mass spectrometry (IM-MS) we characterise changes in the conformational landscape and stability of the protein kinase Aurora A (Aur A) driven by binding of the physiological activator TPX2 or small molecule inhibition. Aided by molecular modelling, we establish three major conformations the relative abundance of which was dependent on Aur A activation status: one highly-populated compact conformer similar to that observed in most crystal structures, a second highly-populated conformer possessing a more open structure infrequently found in crystal structures, and an additional low-abundance conformer not currently represented in the protein databank. Notably, inhibitor binding induces more compact configurations of Aur A, as adopted by the unbound enzyme, with both IM-MS and modelling revealing inhibitor-mediated stabilisation of active Aur A.

Item Type:	Article
Uncontrolled Keywords:	Humans, Protein Conformation, Models, Molecular, Mass Spectrometry, Protein Stability, Aurora Kinase A, Ion Mobility Spectrometry
Divisions:	Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Systems, Molecular and Integrative Biology Faculty of Health and Life Sciences > Tech, Infrastructure and Environmental Directorate
Depositing User:	Symplectic Admin
Date Deposited:	18 May 2022 09:28
Last Modified:	18 Jan 2023 21:01
DOI:	10.1021/jasms.1c00271
Open Access URL:	https://doi.org/10.1021/jasms.1c00271
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3155034