Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain

In response to light, as part of a two-component system, the Brucella blue light-activated histidine kinase (LOV-HK) increases its autophosphorylation, modulating the virulence of this microorganism. The Brucella histidine kinase (HK) domain belongs to the HWE family, for which there is no structura...

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Autores principales:	Rinaldi, Jimena Julieta, Cerutti, María Laura, Berguer, Paula M., Estrin, Dario Ariel, Martí, Marcelo Adrián, Klinke, Sebastián
Publicado:	2016
Materias:	autophosphorylation mechanism molecular dynamics simulations signal transduction two-component system X-ray crystallography dimer homodimer protein histidine kinase protein kinase protein-histidine kinase Article autophosphorylation blue light Brucella conformation cross linking crystal structure crystallography dimerization enzyme activity enzyme structure molecular dynamics mutational analysis nonhuman priority journal protein domain protein family chemistry dna mutational analysis enzymology genetics metabolism phosphorylation protein conformation protein multimerization protein processing X ray crystallography Crystallography, X-Ray DNA Mutational Analysis Molecular Dynamics Simulation Phosphorylation Protein Conformation Protein Kinases Protein Multimerization Protein Processing, Post-Translational
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00222836_v428_n6_p1165_Rinaldi http://hdl.handle.net/20.500.12110/paper_00222836_v428_n6_p1165_Rinaldi
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00222836_v428_n6_p1165_Rinaldi
record_format	dspace
spelling	paper:paper_00222836_v428_n6_p1165_Rinaldi2023-06-08T14:48:46Z Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain Rinaldi, Jimena Julieta Cerutti, María Laura Berguer, Paula M. Estrin, Dario Ariel Martí, Marcelo Adrián Klinke, Sebastián autophosphorylation mechanism molecular dynamics simulations signal transduction two-component system X-ray crystallography dimer homodimer protein histidine kinase protein kinase protein-histidine kinase Article autophosphorylation blue light Brucella conformation cross linking crystal structure crystallography dimerization enzyme activity enzyme structure molecular dynamics mutational analysis nonhuman priority journal protein domain protein family chemistry dna mutational analysis enzymology genetics metabolism phosphorylation protein conformation protein multimerization protein processing X ray crystallography Brucella Crystallography, X-Ray DNA Mutational Analysis Molecular Dynamics Simulation Phosphorylation Protein Conformation Protein Kinases Protein Multimerization Protein Processing, Post-Translational In response to light, as part of a two-component system, the Brucella blue light-activated histidine kinase (LOV-HK) increases its autophosphorylation, modulating the virulence of this microorganism. The Brucella histidine kinase (HK) domain belongs to the HWE family, for which there is no structural information. The HWE family is exclusively present in proteobacteria and usually coupled to a wide diversity of light sensor domains. This work reports the crystal structure of the Brucella HK domain, which presents two different dimeric assemblies in the asymmetric unit: one similar to the already described canonical parallel homodimers (C) and the other, an antiparallel non-canonical (NC) dimer, each with distinct relative subdomain orientations and dimerization interfaces. Contrary to these crystallographic structures and unlike other HKs, in solution, the Brucella HK domain is monomeric and still active, showing an astonishing instability of the dimeric interface. Despite this instability, using cross-linking experiments, we show that the C dimer is the functionally relevant species. Mutational analysis demonstrates that the autophosphorylation activity occurs in cis. The different relative subdomain orientations observed for the NC and C states highlight the large conformational flexibility of the HK domain. Through the analysis of these alternative conformations by means of molecular dynamics simulations, we also propose a catalytic mechanism for Brucella LOV-HK. © 2016 Elsevier Ltd. All rights reserved. Fil:Rinaldi, J. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Cerutti, M.L. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Berguer, P.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Estrín, D.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Martí, M.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Klinke, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2016 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00222836_v428_n6_p1165_Rinaldi http://hdl.handle.net/20.500.12110/paper_00222836_v428_n6_p1165_Rinaldi
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	autophosphorylation mechanism molecular dynamics simulations signal transduction two-component system X-ray crystallography dimer homodimer protein histidine kinase protein kinase protein-histidine kinase Article autophosphorylation blue light Brucella conformation cross linking crystal structure crystallography dimerization enzyme activity enzyme structure molecular dynamics mutational analysis nonhuman priority journal protein domain protein family chemistry dna mutational analysis enzymology genetics metabolism phosphorylation protein conformation protein multimerization protein processing X ray crystallography Brucella Crystallography, X-Ray DNA Mutational Analysis Molecular Dynamics Simulation Phosphorylation Protein Conformation Protein Kinases Protein Multimerization Protein Processing, Post-Translational
spellingShingle	autophosphorylation mechanism molecular dynamics simulations signal transduction two-component system X-ray crystallography dimer homodimer protein histidine kinase protein kinase protein-histidine kinase Article autophosphorylation blue light Brucella conformation cross linking crystal structure crystallography dimerization enzyme activity enzyme structure molecular dynamics mutational analysis nonhuman priority journal protein domain protein family chemistry dna mutational analysis enzymology genetics metabolism phosphorylation protein conformation protein multimerization protein processing X ray crystallography Brucella Crystallography, X-Ray DNA Mutational Analysis Molecular Dynamics Simulation Phosphorylation Protein Conformation Protein Kinases Protein Multimerization Protein Processing, Post-Translational Rinaldi, Jimena Julieta Cerutti, María Laura Berguer, Paula M. Estrin, Dario Ariel Martí, Marcelo Adrián Klinke, Sebastián Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain
topic_facet	autophosphorylation mechanism molecular dynamics simulations signal transduction two-component system X-ray crystallography dimer homodimer protein histidine kinase protein kinase protein-histidine kinase Article autophosphorylation blue light Brucella conformation cross linking crystal structure crystallography dimerization enzyme activity enzyme structure molecular dynamics mutational analysis nonhuman priority journal protein domain protein family chemistry dna mutational analysis enzymology genetics metabolism phosphorylation protein conformation protein multimerization protein processing X ray crystallography Brucella Crystallography, X-Ray DNA Mutational Analysis Molecular Dynamics Simulation Phosphorylation Protein Conformation Protein Kinases Protein Multimerization Protein Processing, Post-Translational
description	In response to light, as part of a two-component system, the Brucella blue light-activated histidine kinase (LOV-HK) increases its autophosphorylation, modulating the virulence of this microorganism. The Brucella histidine kinase (HK) domain belongs to the HWE family, for which there is no structural information. The HWE family is exclusively present in proteobacteria and usually coupled to a wide diversity of light sensor domains. This work reports the crystal structure of the Brucella HK domain, which presents two different dimeric assemblies in the asymmetric unit: one similar to the already described canonical parallel homodimers (C) and the other, an antiparallel non-canonical (NC) dimer, each with distinct relative subdomain orientations and dimerization interfaces. Contrary to these crystallographic structures and unlike other HKs, in solution, the Brucella HK domain is monomeric and still active, showing an astonishing instability of the dimeric interface. Despite this instability, using cross-linking experiments, we show that the C dimer is the functionally relevant species. Mutational analysis demonstrates that the autophosphorylation activity occurs in cis. The different relative subdomain orientations observed for the NC and C states highlight the large conformational flexibility of the HK domain. Through the analysis of these alternative conformations by means of molecular dynamics simulations, we also propose a catalytic mechanism for Brucella LOV-HK. © 2016 Elsevier Ltd. All rights reserved.
author	Rinaldi, Jimena Julieta Cerutti, María Laura Berguer, Paula M. Estrin, Dario Ariel Martí, Marcelo Adrián Klinke, Sebastián
author_facet	Rinaldi, Jimena Julieta Cerutti, María Laura Berguer, Paula M. Estrin, Dario Ariel Martí, Marcelo Adrián Klinke, Sebastián
author_sort	Rinaldi, Jimena Julieta
title	Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain
title_short	Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain
title_full	Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain
title_fullStr	Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain
title_full_unstemmed	Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain
title_sort	structural insights into the hwe histidine kinase family: the brucella blue light-activated histidine kinase domain
publishDate	2016
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00222836_v428_n6_p1165_Rinaldi http://hdl.handle.net/20.500.12110/paper_00222836_v428_n6_p1165_Rinaldi
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Structural Insights into the HWE Histidine Kinase Family: The Brucella Blue Light-Activated Histidine Kinase Domain

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