Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients

Autores
Cozar, Mónica; Urreizti, Roser; Vilarinho, Laura; Grosso, Carola; Dodelson de Kremer, Raquel; Asteggiano, Carla Gabriela; Dalmau, Jaime; García, Ana María; Vilaseca, María Antonia; Grinberg Vaisman, Daniel Raúl; Balcells, Susana
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Homocystinuria due to CBS deficiency is a rare autosomal recessive disorder characterized by elevated plasma levels of homocysteine (Hcy) and methionine (Met). Here we present the analysis of 22 unrelated patients of different geographical origins, mainly Spanish and Argentinian. Twenty-two different mutations were found, 10 of which were novel. Five new mutations were missense and five were deletions of different sizes, including a 794-bp deletion (c.532-37-736 + 438del794) detected by Southern blot analysis. To assess the pathogenicity of these mutations, seven were expressed heterologously in Escherichia coli and their enzyme activities were assayed in vitro, in the absence and presence of the CBS activators PLP and SAM. The presence of the mutant proteins was confirmed by Western blotting. Mutations p.M173del, p.I278S, p.D281N, and p.D321V showed null activity in all conditions tested, whereas mutations p.49L, p.P200L and p.A446S retained different degrees of activity and response to stimulation. Finally, a minigene strategy allowed us to demonstrate the pathogenicity of an 8-bp intronic deletion, which led to the skipping of exon 6. In general, frameshifting deletions correlated with a more severe phenotype, consistent with the concept that missense mutations may recover enzymatic activity under certain conditions.
Fil: Cozar, Mónica. Universidad de Barcelona; España
Fil: Urreizti, Roser. Universidad de Barcelona; España
Fil: Vilarinho, Laura. Instituto de Genética Médica Jacinto Magalhaes; Portugal
Fil: Grosso, Carola. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Estudios de las Metabolopatías Congénitas; Argentina
Fil: Dodelson de Kremer, Raquel. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Estudios de las Metabolopatías Congénitas; Argentina
Fil: Asteggiano, Carla Gabriela. Gobierno de la Provincia de Córdoba. Ministerio de Salud. Hospital de Niños de la Santísima Trinidad; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Dalmau, Jaime. Hospital Infantil La Fe; España
Fil: García, Ana María. Hospital Infantil La Fe; España
Fil: Vilaseca, María Antonia. Hospital Sant Joan de Deu Barcelona; España
Fil: Grinberg Vaisman, Daniel Raúl. Universidad de Barcelona; España
Fil: Balcells, Susana. Universidad de Barcelona; España
Materia
CBS
HETEROLOGOUS EXPRESSION
HOMOCYSTINURIA
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:11336/189554
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/189554
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patientsCozar, MónicaUrreizti, RoserVilarinho, LauraGrosso, CarolaDodelson de Kremer, RaquelAsteggiano, Carla GabrielaDalmau, JaimeGarcía, Ana MaríaVilaseca, María AntoniaGrinberg Vaisman, Daniel RaúlBalcells, SusanaCBSHETEROLOGOUS EXPRESSIONHOMOCYSTINURIAhttps://purl.org/becyt/ford/3.5https://purl.org/becyt/ford/3Homocystinuria due to CBS deficiency is a rare autosomal recessive disorder characterized by elevated plasma levels of homocysteine (Hcy) and methionine (Met). Here we present the analysis of 22 unrelated patients of different geographical origins, mainly Spanish and Argentinian. Twenty-two different mutations were found, 10 of which were novel. Five new mutations were missense and five were deletions of different sizes, including a 794-bp deletion (c.532-37-736 + 438del794) detected by Southern blot analysis. To assess the pathogenicity of these mutations, seven were expressed heterologously in Escherichia coli and their enzyme activities were assayed in vitro, in the absence and presence of the CBS activators PLP and SAM. The presence of the mutant proteins was confirmed by Western blotting. Mutations p.M173del, p.I278S, p.D281N, and p.D321V showed null activity in all conditions tested, whereas mutations p.49L, p.P200L and p.A446S retained different degrees of activity and response to stimulation. Finally, a minigene strategy allowed us to demonstrate the pathogenicity of an 8-bp intronic deletion, which led to the skipping of exon 6. In general, frameshifting deletions correlated with a more severe phenotype, consistent with the concept that missense mutations may recover enzymatic activity under certain conditions.Fil: Cozar, Mónica. Universidad de Barcelona; EspañaFil: Urreizti, Roser. Universidad de Barcelona; EspañaFil: Vilarinho, Laura. Instituto de Genética Médica Jacinto Magalhaes; PortugalFil: Grosso, Carola. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Estudios de las Metabolopatías Congénitas; ArgentinaFil: Dodelson de Kremer, Raquel. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Estudios de las Metabolopatías Congénitas; ArgentinaFil: Asteggiano, Carla Gabriela. Gobierno de la Provincia de Córdoba. Ministerio de Salud. Hospital de Niños de la Santísima Trinidad; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Dalmau, Jaime. Hospital Infantil La Fe; EspañaFil: García, Ana María. Hospital Infantil La Fe; EspañaFil: Vilaseca, María Antonia. Hospital Sant Joan de Deu Barcelona; EspañaFil: Grinberg Vaisman, Daniel Raúl. Universidad de Barcelona; EspañaFil: Balcells, Susana. Universidad de Barcelona; EspañaWiley-liss, div John Wiley & Sons Inc.2011-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/189554Cozar, Mónica; Urreizti, Roser; Vilarinho, Laura; Grosso, Carola; Dodelson de Kremer, Raquel; et al.; Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients; Wiley-liss, div John Wiley & Sons Inc.; Human Mutation; 32; 7; 1-2011; 835-8421059-77941098-1004CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1002/humu.21514info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:14:21Zoai:ri.conicet.gov.ar:11336/189554instacron: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:34982025-10-22 11:14:21.355CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
title Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
spellingShingle Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
Cozar, Mónica
CBS
HETEROLOGOUS EXPRESSION
HOMOCYSTINURIA
title_short Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
title_full Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
title_fullStr Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
title_full_unstemmed Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
title_sort Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients
dc.creator.none.fl_str_mv Cozar, Mónica
Urreizti, Roser
Vilarinho, Laura
Grosso, Carola
Dodelson de Kremer, Raquel
Asteggiano, Carla Gabriela
Dalmau, Jaime
García, Ana María
Vilaseca, María Antonia
Grinberg Vaisman, Daniel Raúl
Balcells, Susana
author Cozar, Mónica
author_facet Cozar, Mónica
Urreizti, Roser
Vilarinho, Laura
Grosso, Carola
Dodelson de Kremer, Raquel
Asteggiano, Carla Gabriela
Dalmau, Jaime
García, Ana María
Vilaseca, María Antonia
Grinberg Vaisman, Daniel Raúl
Balcells, Susana
author_role author
author2 Urreizti, Roser
Vilarinho, Laura
Grosso, Carola
Dodelson de Kremer, Raquel
Asteggiano, Carla Gabriela
Dalmau, Jaime
García, Ana María
Vilaseca, María Antonia
Grinberg Vaisman, Daniel Raúl
Balcells, Susana
author2_role author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv CBS
HETEROLOGOUS EXPRESSION
HOMOCYSTINURIA
topic CBS
HETEROLOGOUS EXPRESSION
HOMOCYSTINURIA
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.5
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Homocystinuria due to CBS deficiency is a rare autosomal recessive disorder characterized by elevated plasma levels of homocysteine (Hcy) and methionine (Met). Here we present the analysis of 22 unrelated patients of different geographical origins, mainly Spanish and Argentinian. Twenty-two different mutations were found, 10 of which were novel. Five new mutations were missense and five were deletions of different sizes, including a 794-bp deletion (c.532-37-736 + 438del794) detected by Southern blot analysis. To assess the pathogenicity of these mutations, seven were expressed heterologously in Escherichia coli and their enzyme activities were assayed in vitro, in the absence and presence of the CBS activators PLP and SAM. The presence of the mutant proteins was confirmed by Western blotting. Mutations p.M173del, p.I278S, p.D281N, and p.D321V showed null activity in all conditions tested, whereas mutations p.49L, p.P200L and p.A446S retained different degrees of activity and response to stimulation. Finally, a minigene strategy allowed us to demonstrate the pathogenicity of an 8-bp intronic deletion, which led to the skipping of exon 6. In general, frameshifting deletions correlated with a more severe phenotype, consistent with the concept that missense mutations may recover enzymatic activity under certain conditions.
Fil: Cozar, Mónica. Universidad de Barcelona; España
Fil: Urreizti, Roser. Universidad de Barcelona; España
Fil: Vilarinho, Laura. Instituto de Genética Médica Jacinto Magalhaes; Portugal
Fil: Grosso, Carola. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Estudios de las Metabolopatías Congénitas; Argentina
Fil: Dodelson de Kremer, Raquel. Universidad Nacional de Córdoba. Facultad de Medicina. Centro de Estudios de las Metabolopatías Congénitas; Argentina
Fil: Asteggiano, Carla Gabriela. Gobierno de la Provincia de Córdoba. Ministerio de Salud. Hospital de Niños de la Santísima Trinidad; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Dalmau, Jaime. Hospital Infantil La Fe; España
Fil: García, Ana María. Hospital Infantil La Fe; España
Fil: Vilaseca, María Antonia. Hospital Sant Joan de Deu Barcelona; España
Fil: Grinberg Vaisman, Daniel Raúl. Universidad de Barcelona; España
Fil: Balcells, Susana. Universidad de Barcelona; España
description Homocystinuria due to CBS deficiency is a rare autosomal recessive disorder characterized by elevated plasma levels of homocysteine (Hcy) and methionine (Met). Here we present the analysis of 22 unrelated patients of different geographical origins, mainly Spanish and Argentinian. Twenty-two different mutations were found, 10 of which were novel. Five new mutations were missense and five were deletions of different sizes, including a 794-bp deletion (c.532-37-736 + 438del794) detected by Southern blot analysis. To assess the pathogenicity of these mutations, seven were expressed heterologously in Escherichia coli and their enzyme activities were assayed in vitro, in the absence and presence of the CBS activators PLP and SAM. The presence of the mutant proteins was confirmed by Western blotting. Mutations p.M173del, p.I278S, p.D281N, and p.D321V showed null activity in all conditions tested, whereas mutations p.49L, p.P200L and p.A446S retained different degrees of activity and response to stimulation. Finally, a minigene strategy allowed us to demonstrate the pathogenicity of an 8-bp intronic deletion, which led to the skipping of exon 6. In general, frameshifting deletions correlated with a more severe phenotype, consistent with the concept that missense mutations may recover enzymatic activity under certain conditions.
publishDate 2011
dc.date.none.fl_str_mv 2011-01
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/11336/189554
Cozar, Mónica; Urreizti, Roser; Vilarinho, Laura; Grosso, Carola; Dodelson de Kremer, Raquel; et al.; Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients; Wiley-liss, div John Wiley & Sons Inc.; Human Mutation; 32; 7; 1-2011; 835-842
1059-7794
1098-1004
CONICET Digital
CONICET
url http://hdl.handle.net/11336/189554
identifier_str_mv Cozar, Mónica; Urreizti, Roser; Vilarinho, Laura; Grosso, Carola; Dodelson de Kremer, Raquel; et al.; Identification and functional analyses of CBS alleles in Spanish and Argentinian homocystinuric patients; Wiley-liss, div John Wiley & Sons Inc.; Human Mutation; 32; 7; 1-2011; 835-842
1059-7794
1098-1004
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1002/humu.21514
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
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley-liss, div John Wiley & Sons Inc.
publisher.none.fl_str_mv Wiley-liss, div John Wiley & Sons 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 12.982451

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