Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/13532
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dc.contributor.authorHenriques, Cristina
dc.contributor.authorMiller, Megan P.
dc.contributor.authorCatanho, Marcos
dc.contributor.authorCarvalho, Técia Maria Ulisses de
dc.contributor.authorKrieger, Marco Aurélio
dc.contributor.authorProbst, Christian M.
dc.contributor.authorSouza, Wanderley de
dc.contributor.authorDegrave, Wim
dc.contributor.authorAmara, Susan Gaye
dc.date.accessioned2016-04-05T15:14:54Z
dc.date.available2016-04-05T15:14:54Z
dc.date.issued2015
dc.identifier.citationHENRIQUES, 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.
dc.identifier.issn1756-3305
dc.identifier.urihttps://www.arca.fiocruz.br/handle/icict/13532
dc.language.isoeng
dc.publisherBioMed Central
dc.rightsopen access
dc.titleIdentification and functional characterization of a novel arginine/ornithine transporter, a member of a cationic amino acid transporter subfamily in the Trypanosoma cruzi genome
dc.typeArticle
dc.identifier.doi10.1186/s13071-015-0950-y
dc.description.abstractenBackground: 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.
dc.creator.affilliationFundação 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.
dc.creator.affilliationUniversity of Pittsburgh. School of Medicine. Department of Neurobiology. Pittsburgh, PA, USA.
dc.creator.affilliationFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil.
dc.creator.affilliationUniversidade 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
dc.creator.affilliationFundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
dc.creator.affilliationFundação Oswaldo Cruz. Instituto Carlos Chagas. Curitiba, PR, Brasil.
dc.creator.affilliationUniversidade 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.
dc.creator.affilliationFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil.
dc.creator.affilliationNational Institute of Mental Health. Bethesda, MD, USA / University of Pittsburgh. School of Medicine. Departmento of Neurobiology. Pittsburg, PA, USA.
dc.subject.enArginine
dc.subject.enOrnithine
dc.subject.enTransporter
dc.subject.enProtozoan
dc.subject.enTrypanosomatids
dc.subject.enT. cruzi
dc.subject.enParasite
dc.subject.esTransportador
dc.subject.decsArginina
dc.subject.decsOrnitina
dc.subject.decsProtozoários
dc.subject.decsTrypanosoma cruzi
dc.subject.decsParasitos
Appears in Collections:PR - ICC - Artigos de Periódicos
MS - Artigos de Periódicos
IOC - Artigos de Periódicos

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