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Published September 11, 2019 | Version v1
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Data from: Lack of activity of recombinant HIF prolyl hydroxylases (PHDs) on reported non-HIF substrates

Authors/Creators

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.

Notes

LC-MS peptide hydroxylation assay data files: Control 1-30
LC-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_1-30.7z
LC-MS peptide hydroxylation assay data files: Control 31-70
LC-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_31-70.7z
LC-MS peptide hydroxylation assay data files: PHD1 1-30
LC-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_1-30.7z
LC-MS peptide hydroxylation assay data files PHD1 31-70
LC-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_31-70.7z
LC-MS peptide hydroxylation assay data files: PHD2 1-30
LC-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_1-30.7z
LC-MS peptide hydroxylation assay data files: PHD2 31-70
LC-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_31-70.7z
LC-MS peptide hydroxylation assay data files: PHD3 1-30
LC-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_1-30.7z
LC-MS peptide hydroxylation assay data files: PHD3 31-70
LC-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_31-70.7z
Peptide Raw file index
Index of MS runs (referenced to enzyme and substrate).

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Peptide Raw file index.pdf

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Is cited by
10.7554/eLife.46490 (DOI)