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dc.contributor.author
Lobo Rojas, Ángel E.
dc.contributor.author
González Marcano, Eglys B.
dc.contributor.author
Valera Vera, Edward Augusto
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dc.contributor.author
Acosta, Rodolfo Héctor
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Quiñones, Wilfredo A
dc.contributor.author
Burchmore, Richard J. S.
dc.contributor.author
Concepción, Juan L.
dc.date.available
2019-09-11T19:47:03Z
dc.date.issued
2016-10
dc.identifier.citation
Lobo Rojas, Ángel E.; González Marcano, Eglys B.; Valera Vera, Edward Augusto; Acosta, Rodolfo Héctor; Quiñones, Wilfredo A; et al.; Trypanosoma cruzi contains two galactokinases; molecular and biochemical characterization; Elsevier Ireland; Parasitology International; 65; 5; 10-2016; 472-482
dc.identifier.issn
1383-5769
dc.identifier.uri
http://hdl.handle.net/11336/83377
dc.description.abstract
Two different putative galactokinase genes, found in the genome database of Trypanosoma cruzi were cloned and sequenced. Expression of the genes in Escherichia coli resulted for TcGALK-1 in the synthesis of a soluble and active enzyme, and in the case of TcGALK-2 gene a less soluble protein, with predicted molecular masses of 51.9 kDa and 51.3 kDa, respectively. The Km values determined for the recombinant proteins were for galactose 0.108 mM (TcGALK-1) and 0.091 mM (TcGALK-2) and for ATP 0.36 mM (TcGALK-1) and 0.1 mM (TcGALK-2). Substrate inhibition by ATP (Ki 0.414 mM) was only observed for TcGALK-2. Gel-filtration chromatography showed that natural TcGALKs and recombinant TcGALK-1 are monomeric. In agreement with the possession of a type-1 peroxisome-targeting signal by both TcGALKs, they were found to be present inside glycosomes using two different methods of subcellular fractionation in conjunction with mass spectrometry. Both genes are expressed in epimastigote and trypomastigote stages since the respective proteins were immunodetected by western blotting. The T. cruzi galactokinases present their highest (52-47%) sequence identity with their counterpart from Leishmania spp., followed by prokaryotic galactokinases such as those from E. coli and Lactococcus lactis (26-23%). In a phylogenetic analysis, the trypanosomatid galactokinases form a separate cluster, showing an affiliation with bacteria. Epimastigotes of T. cruzi can grow in glucose-depleted LIT-medium supplemented with 20 mM of galactose, suggesting that this hexose, upon phosphorylation by a TcGALK, could be used in the synthesis of UDP-galactose and also as a possible carbon and energy source.
dc.format
application/pdf
dc.language.iso
eng
dc.publisher
Elsevier Ireland
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dc.rights
info:eu-repo/semantics/openAccess
dc.rights.uri
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.subject
Galactokinase
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Galactose Metabolism
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Ghmp Superfamily
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Glycosome
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Kinetic Analysis
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Nucleotide Sugars
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Trypanosoma Cruzi
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Udp-Galactose
dc.subject.classification
Bioquímica y Biología Molecular
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dc.subject.classification
Ciencias Biológicas
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CIENCIAS NATURALES Y EXACTAS
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dc.title
Trypanosoma cruzi contains two galactokinases; molecular and biochemical characterization
dc.type
info:eu-repo/semantics/article
dc.type
info:ar-repo/semantics/artículo
dc.type
info:eu-repo/semantics/publishedVersion
dc.date.updated
2019-08-30T13:56:59Z
dc.journal.volume
65
dc.journal.number
5
dc.journal.pagination
472-482
dc.journal.pais
Irlanda
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dc.journal.ciudad
Amsterdam
dc.description.fil
Fil: Lobo Rojas, Ángel E.. Universidad de los Andes; Venezuela
dc.description.fil
Fil: González Marcano, Eglys B.. Universidad de los Andes; Venezuela
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Fil: Valera Vera, Edward Augusto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina. Universidad de los Andes; Venezuela
dc.description.fil
Fil: Acosta, Rodolfo Héctor. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad de los Andes; Venezuela
dc.description.fil
Fil: Quiñones, Wilfredo A. Universidad de los Andes; Venezuela
dc.description.fil
Fil: Burchmore, Richard J. S.. University of Glasgow; Reino Unido
dc.description.fil
Fil: Concepción, Juan L.. Universidad de los Andes; Venezuela
dc.journal.title
Parasitology International
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dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1383576916301891
dc.relation.alternativeid
info:eu-repo/semantics/altIdentifier/doi/http://dx.doi.org/10.1016/j.parint.2016.06.008
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