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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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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. |
format | Online Article Text |
id | pubmed-3688787 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
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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