title: Structural insights into the folding defects of oncogenic pVHL lead to correction of its function in vitro.
abstract: Loss of function mutations in the von Hippel-Lindau (pVHL) tumor suppressor protein are tumorigenic. In silico analysis of the structure and folding of WT pVHL identified in its core an aromatic tetrahedron, essential for stabilizing the protein. The mutations disrupt the aromatic tetrahedron, leading to misfolding of pVHL. Using biophysical methods we confirmed the in silico predictions, demonstrating that mutant pVHL proteins have lower stability than the WT, distort the core domain and as a result reduce the ability of the protein to bind its target HIF-1α. Using bacterial pVHL-EGFP based assay we screened for osmolytes capable of restoring folding of mutant pVHL. Among them, Arginine was the most effective and was verified by in vitro assays as a potent re-folder of pVHL. This resulted in functional restoration of the mutant proteins to the level of the WT.
authors.affiliation: Department of Molecular Microbiology and Biotechnology, George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel.
authors.forename: Merav D
authors.initials: MD
authors.lastname: Shmueli
authors.forename: Lee
authors.initials: L
authors.lastname: Schnaider
authors.forename: Daniel
authors.initials: D
authors.lastname: Rosenblum
authors.forename: Gal
authors.initials: G
authors.lastname: Herzog
authors.forename: Ehud
authors.initials: E
authors.lastname: Gazit
authors.forename: Daniel
authors.initials: D
authors.lastname: Segal
mesh_terms: D001120:Arginine / Q000737:chemistry; D002292:Carcinoma, Renal Cell / Q000235:genetics*; D004926:Escherichia coli; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000737:chemistry; D007680:Kidney Neoplasms / Q000235:genetics*; D008958:Models, Molecular; D008970:Molecular Weight; D020125:Mutation, Missense; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000737:chemistry; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000737:chemistry* / Q000235:genetics
chemical_list: C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D010446:Peptide Fragments; D001120:Arginine; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human