Diamond Publications - Search Results

Search Again...

These results include publications that have not yet been reviewed and processed by Diamond Light Source. To view only processed publications click here.

You can use the folder icon under Actions to collate papers as required. You can then click on View Folder in the left-hand navigation to review and/or download your folder.

One item found (0 items not approved).

Instruments / Technical Area	Publication Details	Published	Actions
I04-Macromolecular Crystallography I24-Microfocus Macromolecular Crystallography	Nek7 conformational flexibility and inhibitor binding probed through protein engineering of the R-spine Matthew J. Byrne , Nazia Nasir , Christine Basmadjian , Chitra Bhatia , Rory F. Cunnison , Katherine H. Carr , Corine Mas-Droux , Sharon Yeoh , Céline Cano , Richard Bayliss Biochemical Journal 477, 1525 - 1539 DOI: 10.1042/BCJ20200128 Diamond Proposal Number(s): [307, 10369, 19248] Open Access Show Abstract ▼ Abstract: Nek7 is a serine/threonine-protein kinase required for proper spindle formation and cytokinesis. Elevated Nek7 levels have been observed in several cancers, and inhibition of Nek7 might provide a route to the development of cancer therapeutics. To date, no selective and potent Nek7 inhibitors have been identified. Nek7 crystal structures exhibit an improperly formed regulatory-spine (R-spine), characteristic of an inactive kinase. We reasoned that the preference of Nek7 to crystallise in this inactive conformation might hinder attempts to capture Nek7 in complex with Type I inhibitors. Here, we have introduced aromatic residues into the R-spine of Nek7 with the aim to stabilise the active conformation of the kinase through R-spine stacking. The strong R-spine mutant Nek7SRS retained catalytic activity and was crystallised in complex with compound 51, an ATP-competitive inhibitor of Nek2 and Nek7. Subsequently, we obtained the same crystal form for wild-type Nek7WT in apo form and bound to compound 51. The R-spines of the three well-ordered Nek7WT molecules exhibit variable conformations while the R-spines of the Nek7SRS molecules all have the same, partially stacked configuration. Compound 51 bound to Nek2 and Nek7 in similar modes, but differences in the precise orientation of a substituent highlights features that could be exploited in designing inhibitors that are selective for particular Nek family members. Although the SRS mutations are not required to obtain a Nek7–inhibitor structure, we conclude that it is a useful strategy for restraining the conformation of a kinase in order to promote crystallogenesis.	Apr 2020

Instruments / Technical Area

Publication Details

Published

Actions

I04-Macromolecular Crystallography
I24-Microfocus Macromolecular Crystallography

Nek7 conformational flexibility and inhibitor binding probed through protein engineering of the R-spine

Matthew J. Byrne , Nazia Nasir , Christine Basmadjian , Chitra Bhatia , Rory F. Cunnison , Katherine H. Carr , Corine Mas-Droux , Sharon Yeoh , Céline Cano , Richard Bayliss

Biochemical Journal 477, 1525 - 1539

DOI: 10.1042/BCJ20200128

Diamond Proposal Number(s): [307, 10369, 19248]