Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates
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...
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2019-09-01
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| Online Access: | https://elifesciences.org/articles/46490 |
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doaj-70d88ea4f82c4443ab414533281020322021-05-05T17:54:34ZengeLife Sciences Publications LtdeLife2050-084X2019-09-01810.7554/eLife.46490Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substratesMatthew E Cockman0https://orcid.org/0000-0002-3310-4821Kerstin Lippl1Ya-Min Tian2Hamish B Pegg3William D Figg Jnr4https://orcid.org/0000-0002-5875-2606Martine I Abboud5https://orcid.org/0000-0003-2141-5988Raphael Heilig6Roman Fischer7https://orcid.org/0000-0002-9715-5951Johanna Myllyharju8https://orcid.org/0000-0001-7772-1250Christopher J Schofield9https://orcid.org/0000-0002-0290-6565Peter J Ratcliffe10https://orcid.org/0000-0002-2853-806XThe Francis Crick Institute, London, United KingdomChemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United KingdomLudwig Institute for Cancer Research, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United KingdomThe Francis Crick Institute, London, United KingdomChemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United KingdomChemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United KingdomTarget Discovery Institute, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United KingdomTarget Discovery Institute, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United KingdomOulu Center for Cell-Matrix Research, Biocenter Oulu and Faculty of Biochemistry and Molecular Medicine, University of Oulu, Oulu, FinlandChemistry Research Laboratory, Department of Chemistry, University of Oxford, Oxford, United KingdomThe Francis Crick Institute, London, United Kingdom; Ludwig Institute for Cancer Research, Nuffield Department of Clinical Medicine, University of Oxford, Oxford, United KingdomHuman 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.https://elifesciences.org/articles/46490hydroxylationprolyl hydroxylaseoxygenase |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Matthew E Cockman Kerstin Lippl Ya-Min Tian Hamish B Pegg William D Figg Jnr Martine I Abboud Raphael Heilig Roman Fischer Johanna Myllyharju Christopher J Schofield Peter J Ratcliffe |
| spellingShingle |
Matthew E Cockman Kerstin Lippl Ya-Min Tian Hamish B Pegg William D Figg Jnr Martine I Abboud Raphael Heilig Roman Fischer Johanna Myllyharju Christopher J Schofield Peter J Ratcliffe Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates eLife hydroxylation prolyl hydroxylase oxygenase |
| author_facet |
Matthew E Cockman Kerstin Lippl Ya-Min Tian Hamish B Pegg William D Figg Jnr Martine I Abboud Raphael Heilig Roman Fischer Johanna Myllyharju Christopher J Schofield Peter J Ratcliffe |
| author_sort |
Matthew E Cockman |
| title |
Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates |
| title_short |
Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates |
| title_full |
Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates |
| title_fullStr |
Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates |
| title_full_unstemmed |
Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates |
| title_sort |
lack of activity of recombinant hif prolyl hydroxylases (phds) on reported non-hif substrates |
| publisher |
eLife Sciences Publications Ltd |
| series |
eLife |
| issn |
2050-084X |
| publishDate |
2019-09-01 |
| description |
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. |
| topic |
hydroxylation prolyl hydroxylase oxygenase |
| url |
https://elifesciences.org/articles/46490 |
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| _version_ |
1721458988700139520 |