Hypoxanthine-guanine phosphoribosyltransferase from Mycobacterium tuberculosis H37Rv: Cloning, expression, and biochemical characterization

Página do item simplificado
dc.contributor.authorBiazus, Gisele
dc.contributor.authorSchneider, Cristopher Z.
dc.contributor.authorPalma, Mario Sergio [UNESP]
dc.contributor.authorBasso, Luiz A.
dc.contributor.authorSantos, Diogenes S.
dc.contributor.institutionPontifícia Universidade Católica do Rio Grande do Sul (PUCRS)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-05-20T13:55:20Z
dc.date.available2014-05-20T13:55:20Z
dc.date.issued2009-08-01
dc.description.abstractHuman tuberculosis (TB) is a major cause of morbidity and mortality worldwide, especially in poor and developing countries. Moreover, the emergence of Mycobacterium tuberculosis strains resistant to first- and second-line anti-TB drugs raises the prospect of virtually incurable TB. Enzymes of the purine phosphoribosyltransferase (PRTase) family are components of purine salvage pathway and have been proposed as drug targets for the development of chemotherapeutic agents against infective and parasitic diseases. The PRTase-catalyzed chemical reaction involves the ribophosphorylation in one step of purine bases (adenine, guanine, hypoxanthine, or xanthine) and their analogues to the respective nucleoside 5'-monophosphate and pyrophosphate. Hypoxanthine-guanine phosphoribosyltransferase (HGPRT; EC 2.4.2.8) is a purine salvage pathway enzyme that specifically recycles hypoxanthine and guanine from the medium, which are in turn converted to, respectively, IMP and GMP. Here we report cloning, DNA sequencing, expression in Escherichia coli BL21 (DE3) cells, purification to homogeneity, N-terminal amino acid sequencing, mass spectrometry analysis, and determination of apparent steady-state kinetic parameters for an in silico predicted M. tuberculosis HGPRT enzyme. These data represent an initial step towards future functional and structural studies, and provide a solid foundation on which to base M. tuberculosis HGPRT-encoding gene manipulation experiments to demonstrate its role in the biology of the bacillus. (C) 2009 Elsevier B.V. All rights reserved.en
dc.description.affiliationPontificia Univ Catolica Rio Grande do Sul, CPBMF, INCT TB, BR-90619900 Porto Alegre, RS, Brazil
dc.description.affiliationPontificia Univ Catolica Rio Grande do Sul, Programa Posgrad Biol Celular & Mol, BR-90619900 Porto Alegre, RS, Brazil
dc.description.affiliationUniv Estadual Paulista Rio Claro, Lab Biol Estrutural & Zooquim, Ctr Estudos Insetos Sociais, Dept Biol,Inst Biociencias, BR-13506900 São Paulo, Brazil
dc.description.affiliationUnespUniv Estadual Paulista Rio Claro, Lab Biol Estrutural & Zooquim, Ctr Estudos Insetos Sociais, Dept Biol,Inst Biociencias, BR-13506900 São Paulo, Brazil
dc.description.sponsorshipNational Institute of Science and Technology on Tuberculosis
dc.description.sponsorshipFINER
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipLaboratório Americano de Farmacoterapia (FARMASA)
dc.description.sponsorshipBanco Nacional de Desenvolvimento Econômico e Social (BNDES)
dc.description.sponsorshipIdFINER: 304051/1975-06
dc.description.sponsorshipIdFINER: 520182/99-5
dc.description.sponsorshipIdFINER: 500079/90-0
dc.format.extent185-190
dc.identifierhttp://dx.doi.org/10.1016/j.pep.2009.04.001
dc.identifier.citationProtein Expression and Purification. San Diego: Academic Press Inc. Elsevier B.V., v. 66, n. 2, p. 185-190, 2009.
dc.identifier.doi10.1016/j.pep.2009.04.001
dc.identifier.issn1046-5928
dc.identifier.lattes2901888624506535
dc.identifier.urihttp://hdl.handle.net/11449/19804
dc.identifier.wosWOS:000266394800010
dc.language.isoeng
dc.publisherAcademic Press Inc. Elsevier B.V.
dc.relation.ispartofProtein Expression and Purification
dc.relation.ispartofjcr1.338
dc.relation.ispartofsjr0,648
dc.rights.accessRightsAcesso restrito
dc.sourceWeb of Science
dc.subjectMycobacterium tuberculosisen
dc.subjectPurine salvage pathwayen
dc.subjectHypoxanthine-guanine phosphoribosyltransferaseen
dc.subjectHGPRTen
dc.subjectProtein purificationen
dc.subjectKinetic parametersen
dc.titleHypoxanthine-guanine phosphoribosyltransferase from Mycobacterium tuberculosis H37Rv: Cloning, expression, and biochemical characterizationen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderAcademic Press Inc. Elsevier B.V.
unesp.author.lattes2901888624506535
unesp.author.orcid0000-0002-7363-8211[3]
unesp.author.orcid0000-0003-0903-2407[4]
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, Rio Claropt

Arquivos

Licença do Pacote
Agora exibindo 1 - 2 de 2
Nenhuma Miniatura disponível
Nome:
license.txt
Tamanho:
1.71 KB
Formato:
Item-specific license agreed upon to submission
Descrição:
Nenhuma Miniatura disponível
Nome:
license.txt
Tamanho:
1.71 KB
Formato:
Item-specific license agreed upon to submission
Descrição:

Coleções

Artigos - Biologia - IBRC