Issue 6, 2023

Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5

Abstract

Jumonji-C (JmjC) domain-containing protein 5 (JMJD5) is a human 2-oxoglutarate (2OG) and Fe(II)-dependent oxygenase which catalyses the post-translational C3 hydroxylation of arginyl-residues and which is linked to the circadian rhythm and to cancer biology through as yet unidentified mechanisms. We report robust solid phase extraction coupled to mass spectrometry (SPE-MS)-based JMJD5 assays which enable kinetic and high-throughput inhibition studies. The kinetic studies reveal that some synthetic 2OG derivatives, notably including a 2OG derivative with a cyclic carbon backbone (i.e. (1R)-3-(carboxycarbonyl)cyclopentane-1-carboxylic acid), are efficient alternative cosubstrates of JMJD5 and of factor inhibiting hypoxia-inducible transcription factor HIF-α (FIH), but not of the Jumonji-C (JmjC) histone Nε-methyl lysine demethylase KDM4E, apparently reflecting the closer structural similarity of JMJD5 and FIH. The JMJD5 inhibition assays were validated by investigating the effect of reported 2OG oxygenase inhibitors on JMJD5 catalysis; the results reveal that broad-spectrum 2OG oxygenase inhibitors are also efficient JMJD5 inhibitors (e.g. N-oxalylglycine, pyridine-2,4-dicarboxylic acid, ebselen) whereas most 2OG oxygenase inhibitors that are in clinical use (e.g. roxadustat) do not inhibit JMJD5. The SPE-MS assays will help enable the development of efficient and selective JMJD5 inhibitors for investigating the biochemical functions of JMJD5 in cellular studies.

Graphical abstract: Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5

Supplementary files

Article information

Article type
Paper
Submitted
16 dek 2022
Accepted
19 mar 2023
First published
20 mar 2023
This article is Open Access
Creative Commons BY license

RSC Chem. Biol., 2023,4, 399-413

Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5

A. Tumber, E. Salah, L. Brewitz, T. P. Corner and C. J. Schofield, RSC Chem. Biol., 2023, 4, 399 DOI: 10.1039/D2CB00249C

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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