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Mapping the interaction between eukaryotic initiation factor 4A (eIF4A) and the inhibitor hippuristanol using carbene footprinting and mass spectrometry

Lloyd, James R.; Hogan, Amy; Paschalis, Vasileios; Bellamy-Carter, Jeddidiah; Bottley, Andrew; Seymour, Graham B.; Hayes, Christopher J.; Oldham, Neil J.

Mapping the interaction between eukaryotic initiation factor 4A (eIF4A) and the inhibitor hippuristanol using carbene footprinting and mass spectrometry Thumbnail


Authors

James R. Lloyd

Amy Hogan

Vasileios Paschalis

Jeddidiah Bellamy-Carter

Andrew Bottley

Graham B. Seymour

CHRIS HAYES chris.hayes@nottingham.ac.uk
Professor of Organic Chemistry

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NEIL OLDHAM NEIL.OLDHAM@NOTTINGHAM.AC.UK
Professor of Biomolecular Spectrometry



Abstract

Protein-ligand interactions are central to protein activity and cell functionality. Improved knowledge of these relationships greatly benefits our understanding of key biological processes and aids in rational drug design towards the treatment of clinically relevant diseases. Carbene footprinting is a recently developed mass spectrometry-based chemical labelling technique that provides valuable information relating to protein-ligand interactions, such as the mapping of binding sites and associated conformational change. Here, we show the application of carbene footprinting to the interaction between eIF4A helicase and a natural product inhibitor, hippuristanol, found in the coral Isis hippuris. Upon addition of hippuristanol we identified reduced carbene labelling (masking) in regions of eIF4A previously implicated in ligand binding. Additionally, we detected hippuristanol-associated increased carbene labelling (unmasking) around the flexible hinge region of eIF4A, indicating ligand-induced conformational change. This work represents further development of the carbene footprinting technique and demonstrates its potential in characterising medicinally relevant protein-ligand interactions.

Citation

Lloyd, J. R., Hogan, A., Paschalis, V., Bellamy-Carter, J., Bottley, A., Seymour, G. B., …Oldham, N. J. (2021). Mapping the interaction between eukaryotic initiation factor 4A (eIF4A) and the inhibitor hippuristanol using carbene footprinting and mass spectrometry. Proteomics, 21(21-22), Article 2000288. https://doi.org/10.1002/pmic.202000288

Journal Article Type Article
Acceptance Date May 17, 2021
Online Publication Date May 24, 2021
Publication Date 2021-11
Deposit Date Jun 1, 2021
Publicly Available Date Mar 28, 2024
Journal Proteomics
Print ISSN 1615-9853
Electronic ISSN 1615-9861
Publisher Wiley
Peer Reviewed Peer Reviewed
Volume 21
Issue 21-22
Article Number 2000288
DOI https://doi.org/10.1002/pmic.202000288
Keywords Biochemistry; Molecular Biology
Public URL https://nottingham-repository.worktribe.com/output/5619680
Publisher URL https://analyticalsciencejournals.onlinelibrary.wiley.com/doi/10.1002/pmic.202000288
Additional Information This is the peer reviewed version of the following article: Lloyd, J. R., Hogan, A., Paschalis, V., Bellamy-Carter, J., Bottley, A., Seymour, G. B., Hayes, C. J., & Oldham, N. J. (2021). Mapping the interaction between eukaryotic initiation factor 4A (eIF4A) and the inhibitor hippuristanol using carbene footprinting and mass spectrometry. Proteomics. e2000288, which has been published in final form at https://doi.org/10.1002/pmic.202000288. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.

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