A Fluorescent Probe Identifies Active Site Ligands of Inositol Pentakisphosphate 2-Kinase

Whitfield, Hayley, Gilmartin, Megan, Baker, Kendall, Riley, Andrew M, Godage, Himali Y, Potter, Barry VL, Hemmings, Andrew M and Brearley, Charles A (2018) A Fluorescent Probe Identifies Active Site Ligands of Inositol Pentakisphosphate 2-Kinase. Journal of Medicinal Chemistry, 61 (19). 8838–8846. ISSN 0022-2623

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Abstract

Inositol pentakisphosphate 2-kinase catalyzes the phosphorylation of the axial 2-OH of myo-inositol 1,3,4,5,6-pentakisphosphate for de novo synthesis of myo-inositol hexakisphosphate. Disruption of inositol pentakisphosphate 2-kinase profoundly influences cellular processes; from nuclear mRNA export and phosphate homeostasis in yeast and plants, to establishment of left-right asymmetry in zebra fish. We elaborate an active site fluorescent probe that allows high throughput screening of Arabidopsis inositol pentakisphosphate 2-kinase. We show that the probe has a binding constant comparable to the Km values of inositol phosphate substrates of this enzyme, and can be used to prospect for novel substrates and inhibitors of inositol phosphate kinases. We identify several micromolar Ki inhibitors and validate this approach by solving the crystal structure of protein in complex with purpurogallin. We additionally solve structures of protein in complexes with epimeric higher inositol phosphates. This probe may find utility in characterization of a wide family of inositol phosphate kinases.

Item Type: Article
Faculty \ School: Faculty of Science > School of Biological Sciences
Faculty of Science > School of Chemistry
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Depositing User: LivePure Connector
Date Deposited: 31 Aug 2018 14:32
Last Modified: 08 Jul 2020 23:55
URI: https://ueaeprints.uea.ac.uk/id/eprint/68178
DOI: 10.1021/acs.jmedchem.8b01022

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