title: Hydroxylation-Dependent Interaction of Substrates to the Von Hippel-Lindau Tumor Suppressor Protein (VHL).
abstract: Oxygen-dependent hydroxylation of critical proline residues, catalyzed by prolyl hydroxylase (PHD1-3) enzymes, is a crucial posttranslational modification (PTM) within the canonical hypoxia-inducible factor (HIF)-centric cellular oxygen-sensing pathway. Alteration of substrates in this way often leads to proteasomal degradation mediated by the von Hippel-Lindau Tumor Suppressor protein (VHL) containing E3-ubiquitin ligase complex known as ECV (Elongins B/C, CUL2, VHL). Here, we outline in vitro protocols to demonstrate the ability of VHL to bind to a prolyl-hydroxylated substrate.
authors: Department of Laboratory Medicine and Pathobiology, University of Toronto, 1 King's College Circle, Toronto, ON, Canada, M5S 1A8.
authors: Pardeep
authors: P
authors: Heir
authors: Department of Laboratory Medicine and Pathobiology, University of Toronto, 1 King's College Circle, Toronto, ON, Canada, M5S 1A8. michael.ohh@utoronto.ca.|Department of Biochemistry, University of Toronto, 1 King's College Circle, Toronto, ON, Canada, M5S 1A8. michael.ohh@utoronto.ca.
authors: Michael
authors: M
authors: Ohh
mesh_terms: D000818:Animals; D006801:Humans; D006900:Hydroxylation; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D066298:In Vitro Techniques; D010100:Oxygen / Q000378:metabolism; D011392:Proline / Q000378:metabolism; D011817:Rabbits; D013379:Substrate Specificity; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D011392:Proline; D051794:Von Hippel-Lindau Tumor Suppressor Protein; D010100:Oxygen
keywords: HIF1α; In vitro binding assay; In vitro hydroxylation; PHD; Proline; VHL