An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys

The human rBAT protein elicits sodium-independent, high affinity obligatory exchange of cystine, dibasic amino acids, and some neutral amino acids in Xenopus oocytes (Chillarón, J., Estévez, R., Mora, C., Wagner, C. A., Suessbrich, H., Lang, F., Gelpí, J. L., Testar, X., Busch, A. E., Zorzano, A., a...

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Autores: Chillarón Chaves, José Julio, Estévez Povedano, Raúl, Samarzija, Ita, Waldegger, Siegfried, Testar, Xavier, Lang, Florian, Zorzano Olarte, Antonio, Busch, Andreas, Palacín Prieto, Manuel
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:1997
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:2445/177125
Acceso en línea:https://hdl.handle.net/2445/177125
Access Level:acceso abierto
Palabra clave:Aminoàcids
Proteïnes portadores
Glicoproteïnes
Metabolisme
Amino acids
Carrier proteins
Glycoproteins
Metabolism
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spelling An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467LysChillarón Chaves, José JulioEstévez Povedano, RaúlSamarzija, ItaWaldegger, SiegfriedTestar, XavierLang, FlorianZorzano Olarte, AntonioBusch, AndreasPalacín Prieto, ManuelAminoàcidsProteïnes portadoresGlicoproteïnesMetabolismeAmino acidsCarrier proteinsGlycoproteinsMetabolismThe human rBAT protein elicits sodium-independent, high affinity obligatory exchange of cystine, dibasic amino acids, and some neutral amino acids in Xenopus oocytes (Chillarón, J., Estévez, R., Mora, C., Wagner, C. A., Suessbrich, H., Lang, F., Gelpí, J. L., Testar, X., Busch, A. E., Zorzano, A., and Palacín, M. (1996) J. Biol. Chem. 271, 17761-17770). Mutations in rBAT have been found to cause cystinuria (Calonge, M. J., Gasparini, P., Chillarón, J., Chillón, M., Galluci, M., Rousaud, F., Zelante, L., Testar, X., Dallapiccola, B., Di Silverio, F., Barceló, P., Estivill, X., Zorzano, A., Nunes, V., and Palacín, M. (1994) Nat. Genet. 6, 420-426). We have performed functional studies with the most common point mutation, M467T, and its relative, M467K, using the oocyte system. The Km and the voltage dependence for transport of the different substrates were the same in both M467T and wild type-injected oocytes. However, the time course of transport was delayed in the M467T mutant: maximal activity was accomplished 3-4 days later than in the wild type. This delay was cRNA dose-dependent: at cRNA levels below 0.5 ng the M467T failed to achieve the wild type transport level. The M467K mutant displayed a normal Km, but the Vmax was between 5 and 35% of the wild type. The amount of rBAT protein was similar in normal and mutant-injected oocytes. In contrast to the wild type, the mutant proteins remained endoglycosidase H-sensitive, suggesting a longer residence time in the endoplasmic reticulum. We quantified the amount of rBAT protein in the plasma membrane by surface labeling with biotin 2 and 6 days after injection. Most of the M467T and M467K protein was located in an intracellular compartment. The converse situation was found in the wild type. Despite the low amount of M467T protein reaching the plasma membrane, the transport activity at 6 days was the same as in the wild type-injected oocytes. The increase in plasma membrane rBAT protein between 2 and 6 days was completely dissociated from the rise in transport activity. These data indicate impaired maturation and transport to the plasma membrane of the M467T and M467K mutant, and suggest that rBAT alone is unable to support the transport function.American Society for Biochemistry and Molecular Biology2021202119972021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion7 p.application/pdfhttps://hdl.handle.net/2445/177125Articles publicats en revistes (Ciències Fisiològiques)reponame:Recercat. Dipósit de la Recerca de Catalunyainstname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)InglésReproducció del document publicat a: https://doi.org/10.1074/jbc.272.14.9543Journal of Biological Chemistry, 1997, vol. 272, num. 14, p. 9543-9549https://doi.org/10.1074/jbc.272.14.9543(c) American Society for Biochemistry and Molecular Biology, 1997info:eu-repo/semantics/openAccessoai:recercat.cat:2445/1771252026-05-29T05:05:01Z
dc.title.none.fl_str_mv An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
title An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
spellingShingle An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
Chillarón Chaves, José Julio
Aminoàcids
Proteïnes portadores
Glicoproteïnes
Metabolisme
Amino acids
Carrier proteins
Glycoproteins
Metabolism
title_short An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
title_full An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
title_fullStr An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
title_full_unstemmed An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
title_sort An intracellular trafficking defect in type-I cystinuria rBAT mutants Met467Thr and Met467Lys
dc.creator.none.fl_str_mv Chillarón Chaves, José Julio
Estévez Povedano, Raúl
Samarzija, Ita
Waldegger, Siegfried
Testar, Xavier
Lang, Florian
Zorzano Olarte, Antonio
Busch, Andreas
Palacín Prieto, Manuel
author Chillarón Chaves, José Julio
author_facet Chillarón Chaves, José Julio
Estévez Povedano, Raúl
Samarzija, Ita
Waldegger, Siegfried
Testar, Xavier
Lang, Florian
Zorzano Olarte, Antonio
Busch, Andreas
Palacín Prieto, Manuel
author_role author
author2 Estévez Povedano, Raúl
Samarzija, Ita
Waldegger, Siegfried
Testar, Xavier
Lang, Florian
Zorzano Olarte, Antonio
Busch, Andreas
Palacín Prieto, Manuel
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Aminoàcids
Proteïnes portadores
Glicoproteïnes
Metabolisme
Amino acids
Carrier proteins
Glycoproteins
Metabolism
topic Aminoàcids
Proteïnes portadores
Glicoproteïnes
Metabolisme
Amino acids
Carrier proteins
Glycoproteins
Metabolism
description The human rBAT protein elicits sodium-independent, high affinity obligatory exchange of cystine, dibasic amino acids, and some neutral amino acids in Xenopus oocytes (Chillarón, J., Estévez, R., Mora, C., Wagner, C. A., Suessbrich, H., Lang, F., Gelpí, J. L., Testar, X., Busch, A. E., Zorzano, A., and Palacín, M. (1996) J. Biol. Chem. 271, 17761-17770). Mutations in rBAT have been found to cause cystinuria (Calonge, M. J., Gasparini, P., Chillarón, J., Chillón, M., Galluci, M., Rousaud, F., Zelante, L., Testar, X., Dallapiccola, B., Di Silverio, F., Barceló, P., Estivill, X., Zorzano, A., Nunes, V., and Palacín, M. (1994) Nat. Genet. 6, 420-426). We have performed functional studies with the most common point mutation, M467T, and its relative, M467K, using the oocyte system. The Km and the voltage dependence for transport of the different substrates were the same in both M467T and wild type-injected oocytes. However, the time course of transport was delayed in the M467T mutant: maximal activity was accomplished 3-4 days later than in the wild type. This delay was cRNA dose-dependent: at cRNA levels below 0.5 ng the M467T failed to achieve the wild type transport level. The M467K mutant displayed a normal Km, but the Vmax was between 5 and 35% of the wild type. The amount of rBAT protein was similar in normal and mutant-injected oocytes. In contrast to the wild type, the mutant proteins remained endoglycosidase H-sensitive, suggesting a longer residence time in the endoplasmic reticulum. We quantified the amount of rBAT protein in the plasma membrane by surface labeling with biotin 2 and 6 days after injection. Most of the M467T and M467K protein was located in an intracellular compartment. The converse situation was found in the wild type. Despite the low amount of M467T protein reaching the plasma membrane, the transport activity at 6 days was the same as in the wild type-injected oocytes. The increase in plasma membrane rBAT protein between 2 and 6 days was completely dissociated from the rise in transport activity. These data indicate impaired maturation and transport to the plasma membrane of the M467T and M467K mutant, and suggest that rBAT alone is unable to support the transport function.
publishDate 1997
dc.date.none.fl_str_mv 1997
2021
2021
2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/177125
url https://hdl.handle.net/2445/177125
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Reproducció del document publicat a: https://doi.org/10.1074/jbc.272.14.9543
Journal of Biological Chemistry, 1997, vol. 272, num. 14, p. 9543-9549
https://doi.org/10.1074/jbc.272.14.9543
dc.rights.none.fl_str_mv (c) American Society for Biochemistry and Molecular Biology, 1997
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) American Society for Biochemistry and Molecular Biology, 1997
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 7 p.
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 Articles publicats en revistes (Ciències Fisiològiques)
reponame:Recercat. Dipósit de la Recerca de Catalunya
instname:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
instname_str Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
reponame_str Recercat. Dipósit de la Recerca de Catalunya
collection Recercat. Dipósit de la Recerca de Catalunya
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.81155

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