Structural basis for modulation of quality control fate in a marginally stable protein
Structural basis for modulation of quality control fate in a marginally stable protein
The human von Hippel-Lindau (VHL) tumor suppressor is a marginally stable protein previously used as a model substrate of eukaryotic refolding and degradation pathways. When expressed in the absence of its cofactors, VHL cannot fold and is quickly degraded by the quality control machinery of the cell. We combined computational methods with in vivo experiments to examine the basis of the misfolding propensity of VHL. By expressing a set of randomly mutated VHL sequences in yeast, we discovered a more stable mutant form. Subsequent modeling suggested the mutation had caused a conformational change affecting cofactor and chaperone interaction, and this hypothesis was then confirmed by additional knockout and overexpression experiments targeting a yeast cofactor homolog. These findings offer a detailed structural basis for the modulation of quality control fate in a model misfolded protein and highlight burial mode modeling as a rapid means to detect functionally important conformational changes in marginally stable globular domains.
Amino Acid Sequence, Amino Acid Substitution, Enzyme Stability, Humans, Models, Molecular, Molecular Sequence Data, Saccharomyces cerevisiae, Von Hippel-Lindau Tumor Suppressor Protein/biosynthesis
1169-78
Brock, Kelly P
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Abraham, Ayelet-chen
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Amen, Triana
388dc540-e819-4d07-8f1e-ee0f3949a54b
Kaganovich, Daniel
ebb13f4e-e925-4aef-88e7-ddc25ef52d8f
England, Jeremy L
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7 July 2015
Brock, Kelly P
d4216bf0-9220-4758-9a10-152b57f014ca
Abraham, Ayelet-chen
9d7bdc49-a883-463d-acce-8635fa88e8b1
Amen, Triana
388dc540-e819-4d07-8f1e-ee0f3949a54b
Kaganovich, Daniel
ebb13f4e-e925-4aef-88e7-ddc25ef52d8f
England, Jeremy L
2f67b12b-9d45-4530-8d48-00034a22e9f9
Brock, Kelly P, Abraham, Ayelet-chen, Amen, Triana, Kaganovich, Daniel and England, Jeremy L
(2015)
Structural basis for modulation of quality control fate in a marginally stable protein.
Structure, 23 (7), .
(doi:10.1016/j.str.2015.04.015).
Abstract
The human von Hippel-Lindau (VHL) tumor suppressor is a marginally stable protein previously used as a model substrate of eukaryotic refolding and degradation pathways. When expressed in the absence of its cofactors, VHL cannot fold and is quickly degraded by the quality control machinery of the cell. We combined computational methods with in vivo experiments to examine the basis of the misfolding propensity of VHL. By expressing a set of randomly mutated VHL sequences in yeast, we discovered a more stable mutant form. Subsequent modeling suggested the mutation had caused a conformational change affecting cofactor and chaperone interaction, and this hypothesis was then confirmed by additional knockout and overexpression experiments targeting a yeast cofactor homolog. These findings offer a detailed structural basis for the modulation of quality control fate in a model misfolded protein and highlight burial mode modeling as a rapid means to detect functionally important conformational changes in marginally stable globular domains.
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Published date: 7 July 2015
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Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.
Keywords:
Amino Acid Sequence, Amino Acid Substitution, Enzyme Stability, Humans, Models, Molecular, Molecular Sequence Data, Saccharomyces cerevisiae, Von Hippel-Lindau Tumor Suppressor Protein/biosynthesis
Identifiers
Local EPrints ID: 475609
URI: http://eprints.soton.ac.uk/id/eprint/475609
ISSN: 0969-2126
PURE UUID: b9239a93-bcb5-4b03-8cfb-251df6997901
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Date deposited: 22 Mar 2023 17:40
Last modified: 17 Mar 2024 04:22
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Author:
Kelly P Brock
Author:
Ayelet-chen Abraham
Author:
Triana Amen
Author:
Daniel Kaganovich
Author:
Jeremy L England
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