Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates
Cockman, Matthew E.; Lippl, Kerstin; Tian, Ya-Min; Pegg, Hamish B.; Figg Jnr, William D.; Abboud, Martine I.; Heilig, Raphael; Fischer, Roman; Myllyharju, Johanna; Schofield, Christopher J.; Ratcliffe, Peter J. (2019-09-10)
eLife 2019;8:e46490, doi: 10.7554/eLife.46490
© 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.
https://creativecommons.org/licenses/by/4.0/
https://urn.fi/URN:NBN:fi-fe2020042422333
Tiivistelmä
Abstract
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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