Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate

Autores
Freeman, Samuel L.; Skafar, Vera; Kwon, Hanna; Fielding, Alistair J.; Moody, Peter C.E.; Martínez, Alejandra; Issoglio, Federico Matías; Inchausti, Lucas; Smircich, Pablo; Zeida, Ari; Piacenza, Lucía; Radi, Rafael; Raven, Emma L.
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The protozoan parasite Trypanosoma cruzi is the causative agent of American trypanosomiasis, otherwise known as Chagas disease. To survive in the host, the T. cruzi parasite needs antioxidant defense systems. One of these is a hybrid heme peroxidase, the T. cruzi ascorbate peroxidase-cytochrome c peroxidase enzyme (TcAPx-CcP). TcAPx-CcP has high sequence identity to members of the class I peroxidase family, notably ascorbate peroxidase (APX) and cytochrome c peroxidase (CcP), as well as a mitochondrial peroxidase from Leishmania major (LmP). The aim of this work was to solve the structure and examine the reactivity of the TcAPx-CcP enzyme. Low temperature electron paramagnetic resonance spectra support the formation of an exchange-coupled [Fe(IV)=O Trp233•+] compound I radical species, analogous to that used in CcP and LmP. We demonstrate that TcAPx-CcP is similar in overall structure to APX and CcP, but there are differences in the substrate-binding regions. Furthermore, the electron transfer pathway from cytochrome c to the heme in CcP and LmP is preserved in the TcAPx-CcP structure. Integration of steady state kinetic experiments, molecular dynamic simulations, and bioinformatic analyses indicates that TcAPx-CcP preferentially oxidizes cytochrome c but is still competent for oxidization of ascorbate. The results reveal that TcAPx-CcP is a credible cytochrome c peroxidase, which can also bind and use ascorbate in host cells, where concentrations are in the millimolar range. Thus, kinetically and functionally TcAPx-CcP can be considered a hybrid peroxidase.
Fil: Freeman, Samuel L.. University of Bristol; Reino Unido
Fil: Skafar, Vera. Universidad de la República; Uruguay
Fil: Kwon, Hanna. University of Leicester; Reino Unido
Fil: Fielding, Alistair J.. Liverpool John Moores University; Reino Unido
Fil: Moody, Peter C.E.. University of Leicester; Reino Unido
Fil: Martínez, Alejandra. Universidad de la República; Uruguay
Fil: Issoglio, Federico Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidade Nova de Lisboa; Portugal
Fil: Inchausti, Lucas. Universidad de la Republica; Uruguay. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay
Fil: Smircich, Pablo. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay. Universidad de la Republica; Uruguay
Fil: Zeida, Ari. Universidad de la Republica; Uruguay
Fil: Piacenza, Lucía. Universidad de la Republica; Uruguay
Fil: Radi, Rafael. Universidad de la Republica; Uruguay
Fil: Raven, Emma L.. University of Bristol; Reino Unido
Materia
ASCORBATE
CHAGAS DISEASE
CYTOCHROME C
HEME
OXIDANTS
PEROXIDASE
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/214995
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/214995
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediateFreeman, Samuel L.Skafar, VeraKwon, HannaFielding, Alistair J.Moody, Peter C.E.Martínez, AlejandraIssoglio, Federico MatíasInchausti, LucasSmircich, PabloZeida, AriPiacenza, LucíaRadi, RafaelRaven, Emma L.ASCORBATECHAGAS DISEASECYTOCHROME CHEMEOXIDANTSPEROXIDASEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The protozoan parasite Trypanosoma cruzi is the causative agent of American trypanosomiasis, otherwise known as Chagas disease. To survive in the host, the T. cruzi parasite needs antioxidant defense systems. One of these is a hybrid heme peroxidase, the T. cruzi ascorbate peroxidase-cytochrome c peroxidase enzyme (TcAPx-CcP). TcAPx-CcP has high sequence identity to members of the class I peroxidase family, notably ascorbate peroxidase (APX) and cytochrome c peroxidase (CcP), as well as a mitochondrial peroxidase from Leishmania major (LmP). The aim of this work was to solve the structure and examine the reactivity of the TcAPx-CcP enzyme. Low temperature electron paramagnetic resonance spectra support the formation of an exchange-coupled [Fe(IV)=O Trp233•+] compound I radical species, analogous to that used in CcP and LmP. We demonstrate that TcAPx-CcP is similar in overall structure to APX and CcP, but there are differences in the substrate-binding regions. Furthermore, the electron transfer pathway from cytochrome c to the heme in CcP and LmP is preserved in the TcAPx-CcP structure. Integration of steady state kinetic experiments, molecular dynamic simulations, and bioinformatic analyses indicates that TcAPx-CcP preferentially oxidizes cytochrome c but is still competent for oxidization of ascorbate. The results reveal that TcAPx-CcP is a credible cytochrome c peroxidase, which can also bind and use ascorbate in host cells, where concentrations are in the millimolar range. Thus, kinetically and functionally TcAPx-CcP can be considered a hybrid peroxidase.Fil: Freeman, Samuel L.. University of Bristol; Reino UnidoFil: Skafar, Vera. Universidad de la República; UruguayFil: Kwon, Hanna. University of Leicester; Reino UnidoFil: Fielding, Alistair J.. Liverpool John Moores University; Reino UnidoFil: Moody, Peter C.E.. University of Leicester; Reino UnidoFil: Martínez, Alejandra. Universidad de la República; UruguayFil: Issoglio, Federico Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidade Nova de Lisboa; PortugalFil: Inchausti, Lucas. Universidad de la Republica; Uruguay. Instituto de Investigaciones Biológicas "Clemente Estable"; UruguayFil: Smircich, Pablo. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay. Universidad de la Republica; UruguayFil: Zeida, Ari. Universidad de la Republica; UruguayFil: Piacenza, Lucía. Universidad de la Republica; UruguayFil: Radi, Rafael. Universidad de la Republica; UruguayFil: Raven, Emma L.. University of Bristol; Reino UnidoAmerican Society for Biochemistry and Molecular Biology2022-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/214995Freeman, Samuel L.; Skafar, Vera; Kwon, Hanna; Fielding, Alistair J.; Moody, Peter C.E.; et al.; Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 298; 8; 8-2022; 1-100021-9258CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbc.2022.102204info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925822006469info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:58:15Zoai:ri.conicet.gov.ar:11336/214995instacron: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:34982025-09-29 09:58:16.206CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
title Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
spellingShingle Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
Freeman, Samuel L.
ASCORBATE
CHAGAS DISEASE
CYTOCHROME C
HEME
OXIDANTS
PEROXIDASE
title_short Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
title_full Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
title_fullStr Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
title_full_unstemmed Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
title_sort Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate
dc.creator.none.fl_str_mv Freeman, Samuel L.
Skafar, Vera
Kwon, Hanna
Fielding, Alistair J.
Moody, Peter C.E.
Martínez, Alejandra
Issoglio, Federico Matías
Inchausti, Lucas
Smircich, Pablo
Zeida, Ari
Piacenza, Lucía
Radi, Rafael
Raven, Emma L.
author Freeman, Samuel L.
author_facet Freeman, Samuel L.
Skafar, Vera
Kwon, Hanna
Fielding, Alistair J.
Moody, Peter C.E.
Martínez, Alejandra
Issoglio, Federico Matías
Inchausti, Lucas
Smircich, Pablo
Zeida, Ari
Piacenza, Lucía
Radi, Rafael
Raven, Emma L.
author_role author
author2 Skafar, Vera
Kwon, Hanna
Fielding, Alistair J.
Moody, Peter C.E.
Martínez, Alejandra
Issoglio, Federico Matías
Inchausti, Lucas
Smircich, Pablo
Zeida, Ari
Piacenza, Lucía
Radi, Rafael
Raven, Emma L.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv ASCORBATE
CHAGAS DISEASE
CYTOCHROME C
HEME
OXIDANTS
PEROXIDASE
topic ASCORBATE
CHAGAS DISEASE
CYTOCHROME C
HEME
OXIDANTS
PEROXIDASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The protozoan parasite Trypanosoma cruzi is the causative agent of American trypanosomiasis, otherwise known as Chagas disease. To survive in the host, the T. cruzi parasite needs antioxidant defense systems. One of these is a hybrid heme peroxidase, the T. cruzi ascorbate peroxidase-cytochrome c peroxidase enzyme (TcAPx-CcP). TcAPx-CcP has high sequence identity to members of the class I peroxidase family, notably ascorbate peroxidase (APX) and cytochrome c peroxidase (CcP), as well as a mitochondrial peroxidase from Leishmania major (LmP). The aim of this work was to solve the structure and examine the reactivity of the TcAPx-CcP enzyme. Low temperature electron paramagnetic resonance spectra support the formation of an exchange-coupled [Fe(IV)=O Trp233•+] compound I radical species, analogous to that used in CcP and LmP. We demonstrate that TcAPx-CcP is similar in overall structure to APX and CcP, but there are differences in the substrate-binding regions. Furthermore, the electron transfer pathway from cytochrome c to the heme in CcP and LmP is preserved in the TcAPx-CcP structure. Integration of steady state kinetic experiments, molecular dynamic simulations, and bioinformatic analyses indicates that TcAPx-CcP preferentially oxidizes cytochrome c but is still competent for oxidization of ascorbate. The results reveal that TcAPx-CcP is a credible cytochrome c peroxidase, which can also bind and use ascorbate in host cells, where concentrations are in the millimolar range. Thus, kinetically and functionally TcAPx-CcP can be considered a hybrid peroxidase.
Fil: Freeman, Samuel L.. University of Bristol; Reino Unido
Fil: Skafar, Vera. Universidad de la República; Uruguay
Fil: Kwon, Hanna. University of Leicester; Reino Unido
Fil: Fielding, Alistair J.. Liverpool John Moores University; Reino Unido
Fil: Moody, Peter C.E.. University of Leicester; Reino Unido
Fil: Martínez, Alejandra. Universidad de la República; Uruguay
Fil: Issoglio, Federico Matías. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química Biológica de la Facultad de Ciencias Exactas y Naturales; Argentina. Universidade Nova de Lisboa; Portugal
Fil: Inchausti, Lucas. Universidad de la Republica; Uruguay. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay
Fil: Smircich, Pablo. Instituto de Investigaciones Biológicas "Clemente Estable"; Uruguay. Universidad de la Republica; Uruguay
Fil: Zeida, Ari. Universidad de la Republica; Uruguay
Fil: Piacenza, Lucía. Universidad de la Republica; Uruguay
Fil: Radi, Rafael. Universidad de la Republica; Uruguay
Fil: Raven, Emma L.. University of Bristol; Reino Unido
description The protozoan parasite Trypanosoma cruzi is the causative agent of American trypanosomiasis, otherwise known as Chagas disease. To survive in the host, the T. cruzi parasite needs antioxidant defense systems. One of these is a hybrid heme peroxidase, the T. cruzi ascorbate peroxidase-cytochrome c peroxidase enzyme (TcAPx-CcP). TcAPx-CcP has high sequence identity to members of the class I peroxidase family, notably ascorbate peroxidase (APX) and cytochrome c peroxidase (CcP), as well as a mitochondrial peroxidase from Leishmania major (LmP). The aim of this work was to solve the structure and examine the reactivity of the TcAPx-CcP enzyme. Low temperature electron paramagnetic resonance spectra support the formation of an exchange-coupled [Fe(IV)=O Trp233•+] compound I radical species, analogous to that used in CcP and LmP. We demonstrate that TcAPx-CcP is similar in overall structure to APX and CcP, but there are differences in the substrate-binding regions. Furthermore, the electron transfer pathway from cytochrome c to the heme in CcP and LmP is preserved in the TcAPx-CcP structure. Integration of steady state kinetic experiments, molecular dynamic simulations, and bioinformatic analyses indicates that TcAPx-CcP preferentially oxidizes cytochrome c but is still competent for oxidization of ascorbate. The results reveal that TcAPx-CcP is a credible cytochrome c peroxidase, which can also bind and use ascorbate in host cells, where concentrations are in the millimolar range. Thus, kinetically and functionally TcAPx-CcP can be considered a hybrid peroxidase.
publishDate 2022
dc.date.none.fl_str_mv 2022-08
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/11336/214995
Freeman, Samuel L.; Skafar, Vera; Kwon, Hanna; Fielding, Alistair J.; Moody, Peter C.E.; et al.; Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 298; 8; 8-2022; 1-10
0021-9258
CONICET Digital
CONICET
url http://hdl.handle.net/11336/214995
identifier_str_mv Freeman, Samuel L.; Skafar, Vera; Kwon, Hanna; Fielding, Alistair J.; Moody, Peter C.E.; et al.; Crystal structure of Trypanosoma cruzi heme peroxidase and characterization of its substrate specificity and compound I intermediate; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 298; 8; 8-2022; 1-10
0021-9258
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbc.2022.102204
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925822006469
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
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 13.070432

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