Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2015 |
| Outros Autores: | , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/13532 |
Resumo: | Fundação Oswaldo Cruz. Campo Grande, MS, Brasil / Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. CENABIO. Núcleo de Biologia Estrutural e Bioimagem. Rio de Janeiro, RJ, Brasil. |
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Henriques, CristinaMiller, Megan P.Catanho, MarcosCarvalho, Técia Maria Ulisses deKrieger, Marco AurélioProbst, Christian M.Souza, Wanderley deDegrave, WimAmara, Susan Gaye2016-04-05T15:14:54Z2016-04-05T15:14:54Z2015HENRIQUES, Cristina et al. Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome. Parasites & Vectors, v. 8, n. 346, p. 1-18, 2015.1756-3305https://www.arca.fiocruz.br/handle/icict/1353210.1186/s13071-015-0950-yengBioMed CentralIdentification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genomeinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Campo Grande, MS, Brasil / Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. CENABIO. Núcleo de Biologia Estrutural e Bioimagem. Rio de Janeiro, RJ, Brasil.University of Pittsburgh. School of Medicine. Department of Neurobiology. Pittsburgh, PA, USA.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, BrasilFundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. CENABIO. Núcleo de Biologia Estrutural e Bioimagem. Rio de Janeiro, RJ, Brasil / Instituto Nacional de Ciência e Tecnologia em Biologia Estrutural e Biomagens - INBEB. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil.National Institute of Mental Health. Bethesda, MD, USA / University of Pittsburgh. School of Medicine. Departmento of Neurobiology. Pittsburg, PA, USA.Background: Trypanosoma cruzi, the etiological agent of Chagas disease, is auxotrophic for arginine. It obtains this amino acid from the host through transporters expressed on the plasma membrane and on the membranes of intracellular compartments. A few cationic amino acid transporters have been characterized at the molecular level, such as the novel intracellular arginine/ornithine transporter, TcCAT1.1, a member of the TcCAT subfamily that is composed of four almost identical open reading frames in the T. cruzi genome. Methods: The functional characterization of the TcCAT1.1 isoform was performed in two heterologous expression systems. TcCAT subfamily expression was evaluated by real-time PCR in polysomal RNA fractions, and the cellular localization of TcCAT1.1 fused to EGFP was performed by confocal and immunoelectron microscopy. Results: In the S. cerevisiae expression system, TcCAT1.1 showed high affinity for arginine (Km = 0.085 ± 0.04 mM) and low affinity for ornithine (Km = 1.7 ± 0.2 mM). Xenopus laevis oocytes expressing TcCAT1.1 showed a 7-fold increase in arginine uptake when they were pre-loaded with arginine, indicating that transport is enhanced by substrates on the trans side of the membrane (trans-stimulation). Oocytes that were pre-loaded with [3 H]-arginine displayed a 16-fold higher efflux of [3 H]-arginine compared with that of the control. Analysis of polysomal RNA fractions demonstrated that the expression of members of the arginine transporter TcCAT subfamily is upregulated under nutritional stress and that this upregulation precedes metacyclogenesis. To investigate the cellular localization of the transporter, EGFP was fused to TcCAT1.1, and fluorescence microscopy and immunocytochemistry revealed the intracellular labeling of vesicles in the anterior region, in a network of tubules and vesicles. Conclusions: TcCAT1.1 is a novel arginine/ornithine transporter, an exchanger expressed in intracellular compartments that is physiologically involved in arginine homeostasis throughout the T. cruzi life cycle. The properties and estimated kinetic parameters of TcCAT1.1 can be extended to other members of the TcCAT subfamily.ArginineOrnithineTransporterProtozoanTrypanosomatidsT. cruziParasiteTransportadorArgininaOrnitinaProtozoáriosTrypanosoma cruziParasitosinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/13532/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALmarcos_catanho_etal_IOC_2015.pdfmarcos_catanho_etal_IOC_2015.pdfapplication/pdf2929685https://www.arca.fiocruz.br/bitstream/icict/13532/2/marcos_catanho_etal_IOC_2015.pdfe211734285e7deec601444ae352c6666MD52TEXTmarcos_catanho_etal_IOC_2015.pdf.txtmarcos_catanho_etal_IOC_2015.pdf.txtExtracted 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| dc.title.pt_BR.fl_str_mv |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| title |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| spellingShingle |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome Henriques, Cristina Arginine Ornithine Transporter Protozoan Trypanosomatids T. cruzi Parasite Transportador Arginina Ornitina Protozoários Trypanosoma cruzi Parasitos |
| title_short |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| title_full |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| title_fullStr |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| title_full_unstemmed |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| title_sort |
Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome |
| author |
Henriques, Cristina |
| author_facet |
Henriques, Cristina Miller, Megan P. Catanho, Marcos Carvalho, Técia Maria Ulisses de Krieger, Marco Aurélio Probst, Christian M. Souza, Wanderley de Degrave, Wim Amara, Susan Gaye |
| author_role |
author |
| author2 |
Miller, Megan P. Catanho, Marcos Carvalho, Técia Maria Ulisses de Krieger, Marco Aurélio Probst, Christian M. Souza, Wanderley de Degrave, Wim Amara, Susan Gaye |
| author2_role |
author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Henriques, Cristina Miller, Megan P. Catanho, Marcos Carvalho, Técia Maria Ulisses de Krieger, Marco Aurélio Probst, Christian M. Souza, Wanderley de Degrave, Wim Amara, Susan Gaye |
| dc.subject.en.pt_BR.fl_str_mv |
Arginine Ornithine Transporter Protozoan Trypanosomatids T. cruzi Parasite |
| topic |
Arginine Ornithine Transporter Protozoan Trypanosomatids T. cruzi Parasite Transportador Arginina Ornitina Protozoários Trypanosoma cruzi Parasitos |
| dc.subject.es.pt_BR.fl_str_mv |
Transportador |
| dc.subject.decs.pt_BR.fl_str_mv |
Arginina Ornitina Protozoários Trypanosoma cruzi Parasitos |
| description |
Fundação Oswaldo Cruz. Campo Grande, MS, Brasil / Universidade Federal do Rio de Janeiro. Centro de Ciências da Saúde. Instituto de Biofísica Carlos Chagas Filho. Laboratório de Ultraestrutura Celular Hertha Meyer. Rio de Janeiro, RJ, Brasil / Universidade Federal do Rio de Janeiro. CENABIO. Núcleo de Biologia Estrutural e Bioimagem. Rio de Janeiro, RJ, Brasil. |
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2015 |
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2015 |
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2016-04-05T15:14:54Z |
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2016-04-05T15:14:54Z |
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HENRIQUES, Cristina et al. Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome. Parasites & Vectors, v. 8, n. 346, p. 1-18, 2015. |
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https://www.arca.fiocruz.br/handle/icict/13532 |
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1756-3305 |
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10.1186/s13071-015-0950-y |
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HENRIQUES, Cristina et al. Identification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome. Parasites & Vectors, v. 8, n. 346, p. 1-18, 2015. 1756-3305 10.1186/s13071-015-0950-y |
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BioMed Central |
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BioMed Central |
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