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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/189554
Ver los metadatos del registro completo
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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 Técnicas2025-09-29T09:45:32Zoai: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:34982025-09-29 09:45:32.679CONICET 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 |
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CONICET Digital (CONICET) |
collection |
CONICET Digital (CONICET) |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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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 |