DOI: 10.5061/dryad.917hb55
title: Data from: Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates

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.
description: LC-MS peptide
 hydroxylation assay data files: Control 1-30LC-MS raw data files
 acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which
 correspond to peptide hydroxylation assays presented in Figure 1 (and
 supplement). See "Peptide Raw file index" pdf for index of MS
 runs (referenced to enzyme and substrate).Control_1-30.7zLC-MS peptide hydroxylation assay data files: Control 31-70LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).Control_31-70.7zLC-MS peptide hydroxylation assay data files: PHD1 1-30LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).PHD1_1-30.7zLC-MS peptide hydroxylation assay data files PHD1 31-70LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).PHD1_31-70.7zLC-MS peptide hydroxylation assay data files: PHD2 1-30LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).PHD2_1-30.7zLC-MS peptide hydroxylation assay data files: PHD2 31-70LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).PHD2_31-70.7zLC-MS peptide hydroxylation assay data files: PHD3 1-30LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).PHD3_1-30.7zLC-MS peptide hydroxylation assay data files: PHD3 31-70LC-MS raw data files acquired on an ACQUITY Xevo G2-S QToF mass spectrometer (Waters) which correspond to peptide hydroxylation assays presented in Figure 1 (and supplement). See "Peptide Raw file index" pdf for index of MS runs (referenced to enzyme and substrate).PHD3_31-70.7zPeptide Raw file indexIndex of MS runs (referenced to enzyme and substrate).