Combined Chemical Modification and Collision Induced Unfolding Using Native Ion Mobility‐Mass Spectrometry Provides Insights into Protein Gas‐Phase Structure

Al‐jabiry, Asia; Palmer, Martin; Langridge, James; Bellamy‐Carter, Jeddidiah; Robinson, David; Oldham, Neil J.

Abstract

Native mass spectrometry is now an important tool in structural biology. Thus, the nature of higher protein structure in the vacuum of the mass spectrometer is an area of significant interest. One of the major goals in the study of gas-phase protein structure is to elucidate the stabilising role of interactions at the level of individual amino acid residues. A strategy combining protein chemical modification together with collision induced unfolding (CIU) was developed and employed to probe the structure of compact protein ions produced by native electrospray ionisation. Tractable chemical modification was used to alter the properties of amino acid residues, and ion mobility-mass spectrometry (IM-MS) utilised to monitor the extent of unfolding as a function of modification. From these data the importance of specific intramolecular interactions for the stability of compact gas-phase protein structure can be inferred. Using this approach, and aided by molecular dynamics simulations, an important stabilising interaction between K6 and H68 in the protein ubiquitin was identified, as was a contact between the N-terminus and E22 in a ubiquitin binding protein UBA2.

Citation

Al‐jabiry, A., Palmer, M., Langridge, J., Bellamy‐Carter, J., Robinson, D., & Oldham, N. J. (2021). Combined Chemical Modification and Collision Induced Unfolding Using Native Ion Mobility‐Mass Spectrometry Provides Insights into Protein Gas‐Phase Structure. Chemistry - A European Journal, 27(55), 13783-13792. https://doi.org/10.1002/chem.202101857

• Journal Article Type: Article
• Acceptance Date: Jul 21, 2021
• Online Publication Date: Jul 21, 2021
• Publication Date: Oct 1, 2021
• Deposit Date: Oct 13, 2021
• Publicly Available Date: Jul 22, 2022
• Journal: Chemistry - A European Journal
• Print ISSN: 0947-6539
• Electronic ISSN: 1521-3765
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 27
• Issue: 55
• Pages: 13783-13792
• DOI: https://doi.org/10.1002/chem.202101857
• Keywords: General Chemistry; Catalysis; Organic Chemistry
• Public URL: https://nottingham-repository.worktribe.com/output/6457283
• Publisher URL: https://chemistry-europe.onlinelibrary.wiley.com/doi/10.1002/chem.202101857
• Additional Information: This is the peer reviewed version of the following article: Al‐jabiry, A., Palmer, M., Langridge, J., Bellamy‐Carter, J., Robinson, D., & Oldham, N. J. (2021). Combined Chemical Modification and Collision Induced Unfolding Using Native Ion Mobility‐Mass Spectrometry Provides Insights into Protein Gas‐Phase Structure. Chemistry - A European Journal, 27(55), 13783-13792, which has been published in final form at https://doi.org/10.1002/chem.202101857. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.