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Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro

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, leadi...

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Autores principales: Shmueli, Merav D., Schnaider, Lee, Rosenblum, Daniel, Herzog, Gal, Gazit, Ehud, Segal, Daniel
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3688787/
https://www.ncbi.nlm.nih.gov/pubmed/23840444
http://dx.doi.org/10.1371/journal.pone.0066333
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author Shmueli, Merav D.
Schnaider, Lee
Rosenblum, Daniel
Herzog, Gal
Gazit, Ehud
Segal, Daniel
author_facet Shmueli, Merav D.
Schnaider, Lee
Rosenblum, Daniel
Herzog, Gal
Gazit, Ehud
Segal, Daniel
author_sort Shmueli, Merav D.
collection PubMed
description 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.
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spelling pubmed-36887872013-07-09 Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro Shmueli, Merav D. Schnaider, Lee Rosenblum, Daniel Herzog, Gal Gazit, Ehud Segal, Daniel PLoS One Research Article 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. Public Library of Science 2013-06-20 /pmc/articles/PMC3688787/ /pubmed/23840444 http://dx.doi.org/10.1371/journal.pone.0066333 Text en © 2013 Shmueli et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Shmueli, Merav D.
Schnaider, Lee
Rosenblum, Daniel
Herzog, Gal
Gazit, Ehud
Segal, Daniel
Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro
title Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro
title_full Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro
title_fullStr Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro
title_full_unstemmed Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro
title_short Structural Insights into the Folding Defects of Oncogenic pVHL Lead to Correction of Its Function In Vitro
title_sort structural insights into the folding defects of oncogenic pvhl lead to correction of its function in vitro
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3688787/
https://www.ncbi.nlm.nih.gov/pubmed/23840444
http://dx.doi.org/10.1371/journal.pone.0066333
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