Decoding Protein Gas‐Phase Stability with Alanine Scanning and Collision‐Induced Unfolding Ion Mobility Mass Spectrometry

Bellamy?Carter, Jeddidiah; O'Grady, Louisa; Passmore, Munro; Jenner, Matthew; Oldham, Neil J.

Abstract

Native mass spectrometry is a widely used tool in structural biology, providing information on protein structure and interactions through preservation of complexes in the gas phase. Herein, the importance of intramolecular non‐covalent interactions in the gas phase has been studied by alanine scanning and collision‐induced unfolding (CIU) ion mobility‐mass spectrometry. Mutation of specific polar and ionic residues on the surface of an acyl carrier protein (ACP) were found to destabilise the compact gas‐phase structure with mutants E31A, D32A, D41A and D65A being particularly destabilised. Molecular dynamics simulations of the ACP 7+ and 8+ ions showed extended intramolecular interactions, resulting from sidechain collapse of polar surface residues, which were confined to the gas phase and consistent with the CIU data. These findings provide evidence for the importance of specific ionic residues, and their interactions, in the maintenance of compact protein gas‐phase structure.

Citation

Bellamy‐Carter, J., O'Grady, L., Passmore, M., Jenner, M., & Oldham, N. J. (2021). Decoding Protein Gas‐Phase Stability with Alanine Scanning and Collision‐Induced Unfolding Ion Mobility Mass Spectrometry. Analysis & Sensing, 1(1), 63-69. https://doi.org/10.1002/anse.202000019

• Journal Article Type: Article
• Acceptance Date: Oct 23, 2020
• Online Publication Date: Oct 23, 2020
• Publication Date: Feb 1, 2021
• Deposit Date: Jan 2, 2021
• Publicly Available Date: Oct 24, 2021
• Journal: Analysis & Sensing
• Print ISSN: 2629-2742
• Electronic ISSN: 2629-2742
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 1
• Issue: 1
• Pages: 63-69
• DOI: https://doi.org/10.1002/anse.202000019
• Keywords: gas-phase protein structure, ion mobility mass spectrometry, molecular dynamics, noncovalent interactions, protein expression
• Public URL: https://nottingham-repository.worktribe.com/output/5184153
• Publisher URL: https://chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/anse.202000019
• Additional Information: This is the peer reviewed version of the following article: Bellamy‐Carter, J., O'Grady, L., Passmore, M., Jenner, M. and Oldham, N.J. (2021), Decoding Protein Gas‐Phase Stability with Alanine Scanning and Collision‐Induced Unfolding Ion Mobility Mass Spectrometry. Analysis & Sensing, which has been published in final form at https://doi.org/10.1002/anse.202000019. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.