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Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter
BACKGROUND: Death-Associated Protein Kinase (DAPK) is a member of the Ca(2+)/calmodulin regulated serine/threonine protein kinases. Its biological function has been associated with induced cell death, and in vivo use of selective small molecule inhibitors of DAPK catalytic activity has demonstrated...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Public Library of Science
2010
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994711/ https://www.ncbi.nlm.nih.gov/pubmed/21152427 http://dx.doi.org/10.1371/journal.pone.0014120 |
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author | Zimmermann, Michael Atmanene, Cédric Xu, Qingyan Fouillen, Laetitia Van Dorsselaer, Alain Bonnet, Dominique Marsol, Claire Hibert, Marcel Sanglier-Cianferani, Sarah Pigault, Claire McNamara, Laurie K. Watterson, D. Martin Haiech, Jacques Kilhoffer, Marie-Claude |
author_facet | Zimmermann, Michael Atmanene, Cédric Xu, Qingyan Fouillen, Laetitia Van Dorsselaer, Alain Bonnet, Dominique Marsol, Claire Hibert, Marcel Sanglier-Cianferani, Sarah Pigault, Claire McNamara, Laurie K. Watterson, D. Martin Haiech, Jacques Kilhoffer, Marie-Claude |
author_sort | Zimmermann, Michael |
collection | PubMed |
description | BACKGROUND: Death-Associated Protein Kinase (DAPK) is a member of the Ca(2+)/calmodulin regulated serine/threonine protein kinases. Its biological function has been associated with induced cell death, and in vivo use of selective small molecule inhibitors of DAPK catalytic activity has demonstrated that it is a potential therapeutic target for treatment of brain injuries and neurodegenerative diseases. METHODOLOGY/PRINCIPAL FINDINGS: In the in vitro study presented here, we describe the homodimerization of DAPK catalytic domain and the crucial role played by its basic loop structure that is part of the molecular fingerprint of death protein kinases. Nanoelectrospray ionization mass spectrometry of DAPK catalytic domain and a basic loop mutant DAPK protein performed under a variety of conditions was used to detect the monomer-dimer interchange. A chemical biological approach was used to find a fluorescent probe that allowed us to follow the oligomerization state of the protein in solution. CONCLUSIONS/SIGNIFICANCE: The use of this combined biophysical and chemical biology approach facilitated the elucidation of a monomer-dimer equilibrium in which the basic loop plays a key role, as well as an apparent allosteric conformational change reported by the fluorescent probe that is independent of the basic loop structure. |
format | Text |
id | pubmed-2994711 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-29947112010-12-08 Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter Zimmermann, Michael Atmanene, Cédric Xu, Qingyan Fouillen, Laetitia Van Dorsselaer, Alain Bonnet, Dominique Marsol, Claire Hibert, Marcel Sanglier-Cianferani, Sarah Pigault, Claire McNamara, Laurie K. Watterson, D. Martin Haiech, Jacques Kilhoffer, Marie-Claude PLoS One Research Article BACKGROUND: Death-Associated Protein Kinase (DAPK) is a member of the Ca(2+)/calmodulin regulated serine/threonine protein kinases. Its biological function has been associated with induced cell death, and in vivo use of selective small molecule inhibitors of DAPK catalytic activity has demonstrated that it is a potential therapeutic target for treatment of brain injuries and neurodegenerative diseases. METHODOLOGY/PRINCIPAL FINDINGS: In the in vitro study presented here, we describe the homodimerization of DAPK catalytic domain and the crucial role played by its basic loop structure that is part of the molecular fingerprint of death protein kinases. Nanoelectrospray ionization mass spectrometry of DAPK catalytic domain and a basic loop mutant DAPK protein performed under a variety of conditions was used to detect the monomer-dimer interchange. A chemical biological approach was used to find a fluorescent probe that allowed us to follow the oligomerization state of the protein in solution. CONCLUSIONS/SIGNIFICANCE: The use of this combined biophysical and chemical biology approach facilitated the elucidation of a monomer-dimer equilibrium in which the basic loop plays a key role, as well as an apparent allosteric conformational change reported by the fluorescent probe that is independent of the basic loop structure. Public Library of Science 2010-11-30 /pmc/articles/PMC2994711/ /pubmed/21152427 http://dx.doi.org/10.1371/journal.pone.0014120 Text en Zimmermann 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 Zimmermann, Michael Atmanene, Cédric Xu, Qingyan Fouillen, Laetitia Van Dorsselaer, Alain Bonnet, Dominique Marsol, Claire Hibert, Marcel Sanglier-Cianferani, Sarah Pigault, Claire McNamara, Laurie K. Watterson, D. Martin Haiech, Jacques Kilhoffer, Marie-Claude Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter |
title | Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter |
title_full | Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter |
title_fullStr | Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter |
title_full_unstemmed | Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter |
title_short | Homodimerization of the Death-Associated Protein Kinase Catalytic Domain: Development of a New Small Molecule Fluorescent Reporter |
title_sort | homodimerization of the death-associated protein kinase catalytic domain: development of a new small molecule fluorescent reporter |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2994711/ https://www.ncbi.nlm.nih.gov/pubmed/21152427 http://dx.doi.org/10.1371/journal.pone.0014120 |
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