An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2

Autores
Zlocowski, Natacha; Lorenz, Virginia; Bennett, Eric P.; Clausen, Henrik; Nores, Gustavo Alejandro; Irazoqui, Fernando Jose
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Polypeptide GalNAc-transferases (ppGalNAc-Ts) are a family of enzymes that catalyze the initiation of mucintype O -glycosylation. All ppGalNAc-T family members contain a common (QXW) 3 motif, which is present in the R-type lectin group. The acetylation site K521 is part of the QKW motif of β -trefoil in the lectin domain of ppGalNAcT2. We used a combination of acetylation and site-directed mutagenesis approaches to examine the functional role of K521 in ppGalNAc-T2. Binding assays of non-acetylated and acetylated forms of the mutant ppGalNAc-T2 K521Q to various naked and α GalNAc-glycosylated mucin peptides indicated that the degree of interaction of lectin domain with α GalNAc depends on the peptide sequence of mucin. Studies of the inhibitory effect of various carbohydrates on the interactions of ppGalNAc-T2 with MUC1 α GalNAc indicate that point K521Q mutation enhance the carbohydrate specificity of lectin domain for α GalNAc. K521Q mutation resulted in an enzyme activity lower than that of the wildtype ppGalNAc-T2, similar to the acetylation of ppGalNAcT2. We conclude that an acetylation site in the QKW motif of the lectin domain modulates carbohydrate recognition specificity and catalytic activity of ppGalNAc-T2 for partially preglycosylated acceptors and a certain naked peptide. Posttranslational modifications of ppGalNAcTs, such as acetylation, may play key roles in modulating the functions of the R-type lectin domains in cellular homeostasis.
Fil: Zlocowski, Natacha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Fil: Lorenz, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Fil: Bennett, Eric P.. Universidad de Copenhagen; Dinamarca
Fil: Clausen, Henrik. Universidad de Copenhagen; Dinamarca
Fil: Nores, Gustavo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Fil: Irazoqui, Fernando Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Materia
Acetilation
Lectin Domain
Ppgalnac-Transferase
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:11336/24987
Acceder
id CONICETDig_d093d04176e887cd5d16fd3801f81b70
oai_identifier_str oai:ri.conicet.gov.ar:11336/24987
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2Zlocowski, NatachaLorenz, VirginiaBennett, Eric P.Clausen, HenrikNores, Gustavo AlejandroIrazoqui, Fernando JoseAcetilationLectin DomainPpgalnac-Transferasehttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Polypeptide GalNAc-transferases (ppGalNAc-Ts) are a family of enzymes that catalyze the initiation of mucintype O -glycosylation. All ppGalNAc-T family members contain a common (QXW) 3 motif, which is present in the R-type lectin group. The acetylation site K521 is part of the QKW motif of β -trefoil in the lectin domain of ppGalNAcT2. We used a combination of acetylation and site-directed mutagenesis approaches to examine the functional role of K521 in ppGalNAc-T2. Binding assays of non-acetylated and acetylated forms of the mutant ppGalNAc-T2 K521Q to various naked and α GalNAc-glycosylated mucin peptides indicated that the degree of interaction of lectin domain with α GalNAc depends on the peptide sequence of mucin. Studies of the inhibitory effect of various carbohydrates on the interactions of ppGalNAc-T2 with MUC1 α GalNAc indicate that point K521Q mutation enhance the carbohydrate specificity of lectin domain for α GalNAc. K521Q mutation resulted in an enzyme activity lower than that of the wildtype ppGalNAc-T2, similar to the acetylation of ppGalNAcT2. We conclude that an acetylation site in the QKW motif of the lectin domain modulates carbohydrate recognition specificity and catalytic activity of ppGalNAc-T2 for partially preglycosylated acceptors and a certain naked peptide. Posttranslational modifications of ppGalNAcTs, such as acetylation, may play key roles in modulating the functions of the R-type lectin domains in cellular homeostasis.Fil: Zlocowski, Natacha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; ArgentinaFil: Lorenz, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; ArgentinaFil: Bennett, Eric P.. Universidad de Copenhagen; DinamarcaFil: Clausen, Henrik. Universidad de Copenhagen; DinamarcaFil: Nores, Gustavo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; ArgentinaFil: Irazoqui, Fernando Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; ArgentinaDe Gruyter2012-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/24987Zlocowski, Natacha; Lorenz, Virginia; Bennett, Eric P.; Clausen, Henrik; Nores, Gustavo Alejandro; et al.; An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2; De Gruyter; Biological Chemistry; 394; 1; 12-2012; 69-771431-6730CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1515/hsz-2012-0191 info:eu-repo/semantics/altIdentifier/url/https://www.degruyter.com/view/j/bchm.2013.394.issue-1/hsz-2012-0191/hsz-2012-0191.xmlinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:54:09Zoai:ri.conicet.gov.ar:11336/24987instacron: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 09:54:09.343CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
title An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
spellingShingle An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
Zlocowski, Natacha
Acetilation
Lectin Domain
Ppgalnac-Transferase
title_short An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
title_full An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
title_fullStr An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
title_full_unstemmed An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
title_sort An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2
dc.creator.none.fl_str_mv Zlocowski, Natacha
Lorenz, Virginia
Bennett, Eric P.
Clausen, Henrik
Nores, Gustavo Alejandro
Irazoqui, Fernando Jose
author Zlocowski, Natacha
author_facet Zlocowski, Natacha
Lorenz, Virginia
Bennett, Eric P.
Clausen, Henrik
Nores, Gustavo Alejandro
Irazoqui, Fernando Jose
author_role author
author2 Lorenz, Virginia
Bennett, Eric P.
Clausen, Henrik
Nores, Gustavo Alejandro
Irazoqui, Fernando Jose
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Acetilation
Lectin Domain
Ppgalnac-Transferase
topic Acetilation
Lectin Domain
Ppgalnac-Transferase
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Polypeptide GalNAc-transferases (ppGalNAc-Ts) are a family of enzymes that catalyze the initiation of mucintype O -glycosylation. All ppGalNAc-T family members contain a common (QXW) 3 motif, which is present in the R-type lectin group. The acetylation site K521 is part of the QKW motif of β -trefoil in the lectin domain of ppGalNAcT2. We used a combination of acetylation and site-directed mutagenesis approaches to examine the functional role of K521 in ppGalNAc-T2. Binding assays of non-acetylated and acetylated forms of the mutant ppGalNAc-T2 K521Q to various naked and α GalNAc-glycosylated mucin peptides indicated that the degree of interaction of lectin domain with α GalNAc depends on the peptide sequence of mucin. Studies of the inhibitory effect of various carbohydrates on the interactions of ppGalNAc-T2 with MUC1 α GalNAc indicate that point K521Q mutation enhance the carbohydrate specificity of lectin domain for α GalNAc. K521Q mutation resulted in an enzyme activity lower than that of the wildtype ppGalNAc-T2, similar to the acetylation of ppGalNAcT2. We conclude that an acetylation site in the QKW motif of the lectin domain modulates carbohydrate recognition specificity and catalytic activity of ppGalNAc-T2 for partially preglycosylated acceptors and a certain naked peptide. Posttranslational modifications of ppGalNAcTs, such as acetylation, may play key roles in modulating the functions of the R-type lectin domains in cellular homeostasis.
Fil: Zlocowski, Natacha. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Fil: Lorenz, Virginia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Fil: Bennett, Eric P.. Universidad de Copenhagen; Dinamarca
Fil: Clausen, Henrik. Universidad de Copenhagen; Dinamarca
Fil: Nores, Gustavo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
Fil: Irazoqui, Fernando Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones En Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Cs.químicas. Centro de Investigaciones En Química Biológica de Córdoba; Argentina
description Polypeptide GalNAc-transferases (ppGalNAc-Ts) are a family of enzymes that catalyze the initiation of mucintype O -glycosylation. All ppGalNAc-T family members contain a common (QXW) 3 motif, which is present in the R-type lectin group. The acetylation site K521 is part of the QKW motif of β -trefoil in the lectin domain of ppGalNAcT2. We used a combination of acetylation and site-directed mutagenesis approaches to examine the functional role of K521 in ppGalNAc-T2. Binding assays of non-acetylated and acetylated forms of the mutant ppGalNAc-T2 K521Q to various naked and α GalNAc-glycosylated mucin peptides indicated that the degree of interaction of lectin domain with α GalNAc depends on the peptide sequence of mucin. Studies of the inhibitory effect of various carbohydrates on the interactions of ppGalNAc-T2 with MUC1 α GalNAc indicate that point K521Q mutation enhance the carbohydrate specificity of lectin domain for α GalNAc. K521Q mutation resulted in an enzyme activity lower than that of the wildtype ppGalNAc-T2, similar to the acetylation of ppGalNAcT2. We conclude that an acetylation site in the QKW motif of the lectin domain modulates carbohydrate recognition specificity and catalytic activity of ppGalNAc-T2 for partially preglycosylated acceptors and a certain naked peptide. Posttranslational modifications of ppGalNAcTs, such as acetylation, may play key roles in modulating the functions of the R-type lectin domains in cellular homeostasis.
publishDate 2012
dc.date.none.fl_str_mv 2012-12
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/11336/24987
Zlocowski, Natacha; Lorenz, Virginia; Bennett, Eric P.; Clausen, Henrik; Nores, Gustavo Alejandro; et al.; An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2; De Gruyter; Biological Chemistry; 394; 1; 12-2012; 69-77
1431-6730
CONICET Digital
CONICET
url http://hdl.handle.net/11336/24987
identifier_str_mv Zlocowski, Natacha; Lorenz, Virginia; Bennett, Eric P.; Clausen, Henrik; Nores, Gustavo Alejandro; et al.; An acetylation site in lectin domain modulates the biological activity of polypeptide GalNAc-transferase-2; De Gruyter; Biological Chemistry; 394; 1; 12-2012; 69-77
1431-6730
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1515/hsz-2012-0191 
info:eu-repo/semantics/altIdentifier/url/https://www.degruyter.com/view/j/bchm.2013.394.issue-1/hsz-2012-0191/hsz-2012-0191.xml
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
application/pdf
dc.publisher.none.fl_str_mv De Gruyter
publisher.none.fl_str_mv De Gruyter
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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