New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM

Autores
Fadel, Firas; Zhao, Yuguang; Cachau, Raul; Cousido Siah, Alexandra; Ruiz, Francesc X.; Harlos, Karl; Howard, Eduardo Ignacio; Mitschler, Andre; Podjarny, Alberto Daniel
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw ) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities.
Fil: Fadel, Firas. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Zhao, Yuguang. University of Oxford; Reino Unido
Fil: Cachau, Raul. Frederick National Laboratory for Cancer Research; Estados Unidos
Fil: Cousido Siah, Alexandra. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Ruiz, Francesc X.. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Harlos, Karl. University of Oxford; Reino Unido
Fil: Howard, Eduardo Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Física de Líquidos y Sistemas Biológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Física de Líquidos y Sistemas Biológicos; Argentina. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Mitschler, Andre. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Podjarny, Alberto Daniel. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Materia
CRYSTAL STRUCTURE
CHITOTRIOSIDASE
CATALYTIC MECHANISM
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/47887
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/47887
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MMFadel, FirasZhao, YuguangCachau, RaulCousido Siah, AlexandraRuiz, Francesc X.Harlos, KarlHoward, Eduardo IgnacioMitschler, AndrePodjarny, Alberto DanielCRYSTAL STRUCTURECHITOTRIOSIDASECATALYTIC MECHANISMhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw ) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities.Fil: Fadel, Firas. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; FranciaFil: Zhao, Yuguang. University of Oxford; Reino UnidoFil: Cachau, Raul. Frederick National Laboratory for Cancer Research; Estados UnidosFil: Cousido Siah, Alexandra. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; FranciaFil: Ruiz, Francesc X.. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; FranciaFil: Harlos, Karl. University of Oxford; Reino UnidoFil: Howard, Eduardo Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Física de Líquidos y Sistemas Biológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Física de Líquidos y Sistemas Biológicos; Argentina. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; FranciaFil: Mitschler, Andre. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; FranciaFil: Podjarny, Alberto Daniel. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; FranciaWiley Blackwell Publishing, Inc2015-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/47887Fadel, Firas; Zhao, Yuguang; Cachau, Raul; Cousido Siah, Alexandra; Ruiz, Francesc X.; et al.; New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM; Wiley Blackwell Publishing, Inc; Acta Crystallographica Section D-Biological Crystallography; 71; 7; 4-2015; 1455-14700907-4449145-1470CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1107/S139900471500783Xinfo:eu-repo/semantics/altIdentifier/url/http://scripts.iucr.org/cgi-bin/paper?S139900471500783Xinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:45:51Zoai:ri.conicet.gov.ar:11336/47887instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:45:51.98CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
title New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
spellingShingle New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
Fadel, Firas
CRYSTAL STRUCTURE
CHITOTRIOSIDASE
CATALYTIC MECHANISM
title_short New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
title_full New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
title_fullStr New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
title_full_unstemmed New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
title_sort New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM
dc.creator.none.fl_str_mv Fadel, Firas
Zhao, Yuguang
Cachau, Raul
Cousido Siah, Alexandra
Ruiz, Francesc X.
Harlos, Karl
Howard, Eduardo Ignacio
Mitschler, Andre
Podjarny, Alberto Daniel
author Fadel, Firas
author_facet Fadel, Firas
Zhao, Yuguang
Cachau, Raul
Cousido Siah, Alexandra
Ruiz, Francesc X.
Harlos, Karl
Howard, Eduardo Ignacio
Mitschler, Andre
Podjarny, Alberto Daniel
author_role author
author2 Zhao, Yuguang
Cachau, Raul
Cousido Siah, Alexandra
Ruiz, Francesc X.
Harlos, Karl
Howard, Eduardo Ignacio
Mitschler, Andre
Podjarny, Alberto Daniel
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CRYSTAL STRUCTURE
CHITOTRIOSIDASE
CATALYTIC MECHANISM
topic CRYSTAL STRUCTURE
CHITOTRIOSIDASE
CATALYTIC MECHANISM
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw ) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities.
Fil: Fadel, Firas. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Zhao, Yuguang. University of Oxford; Reino Unido
Fil: Cachau, Raul. Frederick National Laboratory for Cancer Research; Estados Unidos
Fil: Cousido Siah, Alexandra. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Ruiz, Francesc X.. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Harlos, Karl. University of Oxford; Reino Unido
Fil: Howard, Eduardo Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Física de Líquidos y Sistemas Biológicos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Física de Líquidos y Sistemas Biológicos; Argentina. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Mitschler, Andre. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
Fil: Podjarny, Alberto Daniel. Centre National de la Recherche Scientifique; Francia. Institut de Génétique et de Biologie Moléculaire et Cellulaire; Francia
description Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw ) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities.
publishDate 2015
dc.date.none.fl_str_mv 2015-04
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/47887
Fadel, Firas; Zhao, Yuguang; Cachau, Raul; Cousido Siah, Alexandra; Ruiz, Francesc X.; et al.; New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM; Wiley Blackwell Publishing, Inc; Acta Crystallographica Section D-Biological Crystallography; 71; 7; 4-2015; 1455-1470
0907-4449
145-1470
CONICET Digital
CONICET
url http://hdl.handle.net/11336/47887
identifier_str_mv Fadel, Firas; Zhao, Yuguang; Cachau, Raul; Cousido Siah, Alexandra; Ruiz, Francesc X.; et al.; New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM; Wiley Blackwell Publishing, Inc; Acta Crystallographica Section D-Biological Crystallography; 71; 7; 4-2015; 1455-1470
0907-4449
145-1470
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1107/S139900471500783X
info:eu-repo/semantics/altIdentifier/url/http://scripts.iucr.org/cgi-bin/paper?S139900471500783X
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
publisher.none.fl_str_mv Wiley Blackwell Publishing, Inc
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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