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Validation of IMS‐MS as a screening tool to identify type II kinase inhibitors of FGFR1 kinase

DOI: 10.1002/rcm.9130 DOI Help

Authors: Helen S. Beeston (University of Leeds) , Tobias Klein (AstraZeneca) , Richard A. Norman (AstraZeneca) , Julie A. Tucker (AstraZeneca) , Malcolm Anderson (AstraZeneca) , Alison E. Ashcroft (University of Leeds) , Geoffrey A. Holdgate (AstraZeneca)
Co-authored by industrial partner: Yes

Type: Journal Paper
Journal: Rapid Communications In Mass Spectrometry

State: Published (Approved)
Published: May 2021

Abstract: Rationale: The protein kinase FGFR1 regulates cellular processes in human development. As over-activity of FGFR1 is implicated with cancer, effective inhibitors are in demand. Type I inhibitors, which bind to the active form of FGFR1, are less effective than type II inhibitors, which bind to the inactive form. Screening to distinguish between type I and type II inhibitors is required. Methods: X-Ray crystallography was used to indicate whether a range of potential inhibitors bind to the active or inactive FGFR1 kinase conformation. The binding affinity of each ligand to FGFR1 was measured using biochemical methods. ESI-IMS-MS in conjunction with collision-induced protein unfolding generated a conformational profile of each FGFR1-ligand complex. The results indicate that the protein’s conformational profile depends on whether the inhibitor is type I or type II. Results: X-Ray crystallography confirmed which of the kinase inhibitors bind to the active or inactive form of FGFR1 kinase. Collision-induced unfolding combined with ESI-IMS-MS showed distinct differences in the FGFR1 folding landscape for type I and type II inhibitors. Biochemical studies indicated a similar range of FGFR1 affinities for both types of inhibitors, thus providing confidence that the conformational variations detected using ESI-IMS-MS can be interpretated unequivocally and that this is an effective screening method. Conclusion: A robust ESI-IMS-MS method has been implemented to distinguish between the binding mode of type I and type II inhibitors by monitoring the conformational unfolding profile of FGFR1. This rapid method requires low sample concentrations and could be used as a high-throughput screening technique for the characterisation of novel kinase inhibitors.

Journal Keywords: FGFR1 kinase; screening type I/type II inhibitors; ion mobility spectrometry; mass spectrometry; collision induced unfolding

Subject Areas: Biology and Bio-materials, Chemistry, Medicine


Instruments: I02-Macromolecular Crystallography

Added On: 01/06/2021 08:25

Discipline Tags:

Health & Wellbeing Biochemistry Chemistry Structural biology Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)