Dynamic Equilibrium of the Aurora A Kinase Activation Loop Revealed by Single-Molecule Spectroscopy.
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ICR Authors
Authors
Gilburt, JAH
Sarkar, H
Sheldrake, P
Blagg, J
Ying, L
Dodson, CA
Sarkar, H
Sheldrake, P
Blagg, J
Ying, L
Dodson, CA
Document Type
Journal Article
Date
2017-09
Date Accepted
Abstract
The conformation of the activation loop (T-loop) of protein kinases underlies enzymatic activity and influences the binding of small-molecule inhibitors. By using single-molecule fluorescence spectroscopy, we have determined that phosphorylated Aurora A kinase is in dynamic equilibrium between a DFG-in-like active T-loop conformation and a DFG-out-like inactive conformation, and have measured the rate constants of interconversion. Addition of the Aurora A activating protein TPX2 shifts the equilibrium towards an active T-loop conformation whereas addition of the inhibitors MLN8054 and CD532 favors an inactive T-loop. We show that Aurora A binds TPX2 and MLN8054 simultaneously and provide a new model for kinase conformational behavior. Our approach will enable conformation-specific effects to be integrated into inhibitor discovery across the kinome, and we outline some immediate consequences for structure-based drug discovery.
Citation
Angewandte Chemie (International ed. in English), 2017, 56 (38), pp. 11409 - 11414
Source Title
Publisher
ISSN
1433-7851
eISSN
1521-3773
Collections
Research Team
Medicinal Chemistry 1