Photoaffinity labelling displacement assay using multiple recombinant protein domains

Fallon, David J. and Phillipou, Alex and Schofield, Christopher J. and House, David and Tomkinson, Nicholas C. O. and Bush, Jacob T. (2023) Photoaffinity labelling displacement assay using multiple recombinant protein domains. Biochemical Journal, 480 (15). pp. 1183-1197. ISSN 0264-6021 (https://doi.org/10.1042/bcj20230129)

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Abstract

The development and optimisation of a photoaffinity labelling (PAL) displacement assay is presented, where a highly efficient PAL probe was used to report on the relative binding affinities of compounds to specific binding sites in multiple recombinant protein domains in tandem. The N- and C-terminal bromodomains of BRD4 were used as example target proteins. A test set of 264 compounds annotated with activity against the bromodomain and extra-terminal domain (BET) family in ChEMBL were used to benchmark the assay. The pIC50 values obtained from the assay correlated well with orthogonal TR‑FRET data, highlighting the potential of this highly accessible PAL biochemical screening platform.

ORCID iDs

Fallon, David J. ORCID logoORCID: https://orcid.org/0000-0003-4310-219X, Phillipou, Alex, Schofield, Christopher J., House, David, Tomkinson, Nicholas C. O. ORCID logoORCID: https://orcid.org/0000-0002-5509-0133 and Bush, Jacob T.;
  • Item type:Article
    ID code:86193
    Dates:
    Date
    Event
    31 August 2023
    Published
    4 July 2023
    Published Online
    3 July 2023
    Accepted
    Subjects:Science > Microbiology
    Department:Faculty of Science > Pure and Applied Chemistry
    Depositing user: Pure Administrator
    Date deposited:20 Jul 2023 10:01
    Last modified:11 Nov 2024 14:00
    URI:https://strathprints.strath.ac.uk/id/eprint/86193
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