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Allosteric inhibition of Aurora-A kinase by a synthetic vNAR domain

DOI: 10.1098/rsob.160089 DOI Help

Authors: Selena G. Burgess (University of Leeds) , Arkadiusz Oleksy (Centre for Therapeutics Discovery, MRC Technology) , Tommaso Cavazza (Cell and Developmental Biology program, Centre for Genomic Regulation (CRG) and UPF) , Mark W. Richards (University of Leeds) , Isabelle Vernos (Cell and Developmental Biology program, Centre for Genomic Regulation (CRG) and UPF) , David Matthews (Centre for Therapeutics Discovery, MRC Technology) , Richard Bayliss (University of Leeds; University of Leicester)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Open Biology , VOL 6

State: Published (Approved)
Published: July 2016
Diamond Proposal Number(s): 10369

Open Access Open Access

Abstract: The vast majority of clinically approved protein kinase inhibitors target the ATP-binding pocket directly. Consequently, many inhibitors have broad selectivity profiles and most have significant off-target effects. Allosteric inhibitors are generally more selective, but are difficult to identify because allosteric binding sites are often unknown or poorly characterized. Aurora-A is activated through binding of TPX2 to an allosteric site on the kinase catalytic domain, and this knowledge could be exploited to generate an inhibitor. Here, we generated an allosteric inhibitor of Aurora-A kinase based on a synthetic, vNAR single domain scaffold, vNAR-D01. Biochemical studies and a crystal structure of the Aurora-A/vNAR-D01 complex show that the vNAR domain overlaps with the TPX2 binding site. In contrast with the binding of TPX2, which stabilizes an active conformation of the kinase, binding of the vNAR domain stabilizes an inactive conformation, in which the αC-helix is distorted, the canonical Lys-Glu salt bridge is broken and the regulatory (R-) spine is disrupted by an additional hydrophobic side chain from the activation loop. These studies illustrate how single domain antibodies can be used to characterize the regulatory mechanisms of kinases and provide a rational basis for structure-guided design of allosteric Aurora-A kinase inhibitors.

Journal Keywords: antibody-assisted drug discovery; structural biology; biochemistry; protein kinase

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

Instruments: I04-1-Macromolecular Crystallography (fixed wavelength)