Mapping the interaction between eukaryotic initiation factor 4A (eIF4A) and the inhibitor hippuristanol using carbene footprinting and mass spectrometry. Issue 21 (6th June 2021)

• Record Type: Journal Article
• Authors: Lloyd, James R.; Hogan, Amy; Paschalis, Vasileios; Bellamy‐Carter, Jeddidiah; Bottley, Andrew; Seymour, Graham B.; Hayes, Christopher J.; Oldham, Neil J.
• Other Names: Cooper Helen J. guestEditor.; Leney Aneika C. guestEditor.
• Is Part Of: Proteomics. Volume 21:Issue 21/22(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-06-06
• Subjects: carbene labelling -- eIF4A -- mass spectrometry -- protein footprinting; Proteins -- Separation -- Periodicals; Bioinformatics -- Periodicals; Proteomics -- Periodicals; Genomes -- Periodicals; Molecular genetics -- Periodicals; 572.605
• DOI: 10.1002/pmic.202000288
• Languages: English
• ISSNs: 1615-9853

Abstract

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.