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

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...

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Autores principales: Tumber, Anthony, Salah, Eidarus, Brewitz, Lennart, Corner, Thomas P., Schofield, Christopher J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: RSC 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10246557/
https://www.ncbi.nlm.nih.gov/pubmed/37292060
http://dx.doi.org/10.1039/d2cb00249c
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author Tumber, Anthony
Salah, Eidarus
Brewitz, Lennart
Corner, Thomas P.
Schofield, Christopher J.
author_facet Tumber, Anthony
Salah, Eidarus
Brewitz, Lennart
Corner, Thomas P.
Schofield, Christopher J.
author_sort Tumber, Anthony
collection PubMed
description 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 N(ε)-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.
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spelling pubmed-102465572023-06-08 Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5 Tumber, Anthony Salah, Eidarus Brewitz, Lennart Corner, Thomas P. Schofield, Christopher J. RSC Chem Biol Chemistry 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 N(ε)-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. RSC 2023-03-20 /pmc/articles/PMC10246557/ /pubmed/37292060 http://dx.doi.org/10.1039/d2cb00249c Text en This journal is © The Royal Society of Chemistry https://creativecommons.org/licenses/by/3.0/
spellingShingle Chemistry
Tumber, Anthony
Salah, Eidarus
Brewitz, Lennart
Corner, Thomas P.
Schofield, Christopher J.
Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5
title Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5
title_full Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5
title_fullStr Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5
title_full_unstemmed Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5
title_short Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5
title_sort kinetic and inhibition studies on human jumonji-c (jmjc) domain-containing protein 5
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10246557/
https://www.ncbi.nlm.nih.gov/pubmed/37292060
http://dx.doi.org/10.1039/d2cb00249c
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