Abstract
The product of the von Hippel-Lindau gene, pVHL, targets the alpha subunits of the heterodimeric transcription factor hypoxia-inducible factor (HIF) for polyubiquitination in the presence of oxygen. The binding of pVHL to HIF is governed by the enzymatic hydroxylation of conserved prolyl residues within peptidic motifs present in the HIFalpha family members. By using a biochemical purification strategy, we have identified a human homolog of Caenorhabditis elegans Egl9 as a HIF prolyl hydroxylase. In addition, we studied the activity of a structurally diverse collection of low molecular weight inhibitors of procollagen prolyl 4-hydroxylase as potential inhibitors of the HIF hydroxylase. A model compound of this series stabilized HIF in a variety of cells, leading to the increased production of its downstream target, vascular endothelial growth factor.
MeSH terms
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Amino Acid Sequence
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Animals
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Base Sequence
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Caenorhabditis elegans Proteins / metabolism
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Cell Line
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Cloning, Molecular
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DNA, Complementary / genetics
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / metabolism*
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Drug Stability
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Enzyme Inhibitors / chemistry
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Enzyme Inhibitors / pharmacology
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Helminth Proteins / metabolism
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Humans
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Hydroxylation
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Hypoxia-Inducible Factor 1
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Hypoxia-Inducible Factor 1, alpha Subunit
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In Vitro Techniques
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Ligases / metabolism
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Molecular Sequence Data
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Nuclear Proteins / genetics
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Nuclear Proteins / metabolism*
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Procollagen-Proline Dioxygenase / antagonists & inhibitors*
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Procollagen-Proline Dioxygenase / isolation & purification*
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Procollagen-Proline Dioxygenase / metabolism
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Rabbits
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Reticulocytes / enzymology
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Transcription Factors*
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Tumor Suppressor Proteins*
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Ubiquitin-Protein Ligases*
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Von Hippel-Lindau Tumor Suppressor Protein
Substances
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Caenorhabditis elegans Proteins
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DNA, Complementary
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DNA-Binding Proteins
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Egl-9 protein, C elegans
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Enzyme Inhibitors
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HIF1A protein, human
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Helminth Proteins
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Hypoxia-Inducible Factor 1
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Hypoxia-Inducible Factor 1, alpha Subunit
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Nuclear Proteins
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Recombinant Fusion Proteins
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Transcription Factors
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Tumor Suppressor Proteins
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Procollagen-Proline Dioxygenase
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Ubiquitin-Protein Ligases
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Von Hippel-Lindau Tumor Suppressor Protein
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Ligases
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VHL protein, human