Cookies on this website
We use cookies to ensure that we give you the best experience on our website. If you click 'Continue' we'll assume that you are happy to receive all cookies and you won't see this message again. Click 'Find out more' for information on how to change your cookie settings.
Skip to main content

Protein kinases are critical therapeutic targets. Pim kinases are implicated in several leukaemias and cancers. Here, we exploit a protein nanopore sensor for Pim kinases that bears a pseudosubstrate peptide attached by an enhanced engineering approach. Analyte binding to the sensor peptide is measured through observation of the modulation of ionic current through a single nanopore. We observed synergistic binding of MgATP and kinase to the sensor, which was used to develop a superior method to evaluate Pim kinase inhibitors featuring label-free determination of inhibition constants. The procedure circumvents many sources of bias or false-positives inherent in current assays. For example, we identified a potent inhibitor missed by differential scanning fluorimetry. The approach is also amenable to implementation on high throughput chips.

Original publication

DOI

10.1002/anie.201503141

Type

Journal article

Journal

Angew Chem Int Ed Engl

Publication Date

06/07/2015

Volume

54

Pages

8154 - 8159

Keywords

drug discovery, inhibitors, protein kinases, screening, single-molecule studies, Drug Discovery, Molecular Structure, Nanopores, Protein Kinase Inhibitors, Protein Kinases, Proto-Oncogene Proteins c-pim-1