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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Arginine / chemistry
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Carcinoma, Renal Cell / genetics*
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Escherichia coli
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Humans
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Hypoxia-Inducible Factor 1, alpha Subunit / chemistry
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Kidney Neoplasms / genetics*
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Models, Molecular
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Molecular Weight
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Mutation, Missense
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Osmolar Concentration
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Peptide Fragments / chemistry
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Protein Binding
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Protein Folding
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Protein Structure, Tertiary
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Von Hippel-Lindau Tumor Suppressor Protein / chemistry*
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Von Hippel-Lindau Tumor Suppressor Protein / genetics
Substances
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HIF1A protein, human
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Hypoxia-Inducible Factor 1, alpha Subunit
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Peptide Fragments
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Arginine
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Von Hippel-Lindau Tumor Suppressor Protein
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VHL protein, human
Grants and funding
This work was supported in part by grants from the Israel Cancer Association (
http://en.cancer.org.il/) and the Cancer Biology Research Center, Tel Aviv University,(
http://www.tau.ac.il/institutes/cbrc/) to DS. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.