University of Oulu

eLife 2019;8:e46490, doi: 10.7554/eLife.46490

Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates

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Author: Cockman, Matthew E.1; Lippl, Kerstin2; Tian, Ya-Min3;
Organizations: 1The Francis Crick Institute, London, United Kingdom
2Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United Kingdom
3Ludwig Institute for Cancer Research, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United Kingdom
4Target Discovery Institute, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United Kingdom
5Oulu Center for Cell-Matrix Research, Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 4 MB)
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Language: English
Published: eLife Sciences Publications, 2019
Publish Date: 2020-04-24


Human and other animal cells deploy three closely related dioxygenases (PHD 1, 2 and 3) to signal oxygen levels by catalysing oxygen regulated prolyl hydroxylation of the transcription factor HIF. The discovery of the HIF prolyl-hydroxylase (PHD) enzymes as oxygen sensors raises a key question as to the existence and nature of non-HIF substrates, potentially transducing other biological responses to hypoxia. Over 20 such substrates are reported. We therefore sought to characterise their reactivity with recombinant PHD enzymes. Unexpectedly, we did not detect prolyl-hydroxylase activity on any reported non-HIF protein or peptide, using conditions supporting robust HIF-α hydroxylation. We cannot exclude PHD-catalysed prolyl hydroxylation occurring under conditions other than those we have examined. However, our findings using recombinant enzymes provide no support for the wide range of non-HIF PHD substrates that have been reported.

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Series: eLife
ISSN: 2050-084X
ISSN-E: 2050-084X
ISSN-L: 2050-084X
Volume: 8
Article number: e46490
DOI: 10.7554/eLife.46490
Type of Publication: A1 Journal article – refereed
Field of Science: 1182 Biochemistry, cell and molecular biology
Funding: This work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001501), the UK Medical Research Council (FC001501), and the Wellcome Trust (FC001501). PJR is also supported by the Ludwig Institute for Cancer Research and the Wellcome Trust (106241/Z/14/Z). CJS, KL, MA, and WDF thank the Wellcome Trust (106244/Z/14/Z), Cancer Research UK, and the British Heart Foundation for funding. KL gratefully acknowledges support via the Newton Abraham D Phil studentship scheme. JM was supported by Academy of Finland project grant 296498, Academy of Finland Center of Excellence 2012–2017 Grant 251314, the S Jusèlius Foundation and the Jane and Aatos Erkko Foundation.
Academy of Finland Grant Number: 296498
Detailed Information: 296498 (Academy of Finland Funding decision)
251314 (Academy of Finland Funding decision)
Copyright information: © Cockman et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited.