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Molecular motion regulates the activity of the Mitochondrial Serine Protease HtrA2
HtrA2 (high-temperature requirement 2) is a human mitochondrial protease that has a role in apoptosis and Parkinson’s disease. The structure of HtrA2 with an intact catalytic triad was determined, revealing a conformational change in the active site loops, involving mainly the regulatory LD loop, wh...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5759095/ https://www.ncbi.nlm.nih.gov/pubmed/29022916 http://dx.doi.org/10.1038/cddis.2017.487 |
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author | Merski, Matthew Moreira, Cátia Abreu, Rui MV Ramos, Maria João Fernandes, Pedro A Martins, L Miguel Pereira, Pedro José Barbosa Macedo-Ribeiro, Sandra |
author_facet | Merski, Matthew Moreira, Cátia Abreu, Rui MV Ramos, Maria João Fernandes, Pedro A Martins, L Miguel Pereira, Pedro José Barbosa Macedo-Ribeiro, Sandra |
author_sort | Merski, Matthew |
collection | PubMed |
description | HtrA2 (high-temperature requirement 2) is a human mitochondrial protease that has a role in apoptosis and Parkinson’s disease. The structure of HtrA2 with an intact catalytic triad was determined, revealing a conformational change in the active site loops, involving mainly the regulatory LD loop, which resulted in burial of the catalytic serine relative to the previously reported structure of the proteolytically inactive mutant. Mutations in the loops surrounding the active site that significantly restricted their mobility, reduced proteolytic activity both in vitro and in cells, suggesting that regulation of HtrA2 activity cannot be explained by a simple transition to an activated conformational state with enhanced active site accessibility. Manipulation of solvent viscosity highlighted an unusual bi-phasic behavior of the enzymatic activity, which together with MD calculations supports the importance of motion in the regulation of the activity of HtrA2. HtrA2 is an unusually thermostable enzyme (T(M)=97.3 °C), a trait often associated with structural rigidity, not dynamic motion. We suggest that this thermostability functions to provide a stable scaffold for the observed loop motions, allowing them a relatively free conformational search within a rather restricted volume. |
format | Online Article Text |
id | pubmed-5759095 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-57590952018-02-14 Molecular motion regulates the activity of the Mitochondrial Serine Protease HtrA2 Merski, Matthew Moreira, Cátia Abreu, Rui MV Ramos, Maria João Fernandes, Pedro A Martins, L Miguel Pereira, Pedro José Barbosa Macedo-Ribeiro, Sandra Cell Death Dis Original Article HtrA2 (high-temperature requirement 2) is a human mitochondrial protease that has a role in apoptosis and Parkinson’s disease. The structure of HtrA2 with an intact catalytic triad was determined, revealing a conformational change in the active site loops, involving mainly the regulatory LD loop, which resulted in burial of the catalytic serine relative to the previously reported structure of the proteolytically inactive mutant. Mutations in the loops surrounding the active site that significantly restricted their mobility, reduced proteolytic activity both in vitro and in cells, suggesting that regulation of HtrA2 activity cannot be explained by a simple transition to an activated conformational state with enhanced active site accessibility. Manipulation of solvent viscosity highlighted an unusual bi-phasic behavior of the enzymatic activity, which together with MD calculations supports the importance of motion in the regulation of the activity of HtrA2. HtrA2 is an unusually thermostable enzyme (T(M)=97.3 °C), a trait often associated with structural rigidity, not dynamic motion. We suggest that this thermostability functions to provide a stable scaffold for the observed loop motions, allowing them a relatively free conformational search within a rather restricted volume. Nature Publishing Group 2017-10 2017-10-12 /pmc/articles/PMC5759095/ /pubmed/29022916 http://dx.doi.org/10.1038/cddis.2017.487 Text en Copyright © 2017 The Author(s) http://creativecommons.org/licenses/by/4.0/ Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Original Article Merski, Matthew Moreira, Cátia Abreu, Rui MV Ramos, Maria João Fernandes, Pedro A Martins, L Miguel Pereira, Pedro José Barbosa Macedo-Ribeiro, Sandra Molecular motion regulates the activity of the Mitochondrial Serine Protease HtrA2 |
title | Molecular motion regulates the activity of the Mitochondrial Serine Protease
HtrA2 |
title_full | Molecular motion regulates the activity of the Mitochondrial Serine Protease
HtrA2 |
title_fullStr | Molecular motion regulates the activity of the Mitochondrial Serine Protease
HtrA2 |
title_full_unstemmed | Molecular motion regulates the activity of the Mitochondrial Serine Protease
HtrA2 |
title_short | Molecular motion regulates the activity of the Mitochondrial Serine Protease
HtrA2 |
title_sort | molecular motion regulates the activity of the mitochondrial serine protease
htra2 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5759095/ https://www.ncbi.nlm.nih.gov/pubmed/29022916 http://dx.doi.org/10.1038/cddis.2017.487 |
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