Biochemical characterization of Neurospora crassa glycogenin (GNN), the self-glucosylating initiator of glycogen synthesis

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dc.contributor.authorde Paula, R. M.
dc.contributor.authorWilson, W. A.
dc.contributor.authorRoach, P. J.
dc.contributor.authorTerenzi, H. F.
dc.contributor.authorBertolini, Maria Celia [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionIndiana University
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.date.accessioned2014-05-20T15:20:19Z
dc.date.available2014-05-20T15:20:19Z
dc.date.issued2005-04-11
dc.description.abstractGlycogenin acts in the initiation step of glycogen biosynthesis by catalyzing a self-glucosylation reaction. In a previous work [de Paula et al., Arch. Biochem. Biophys. 435 (2005) 112-124], we described the isolation of the cDNA gnn, which encodes the protein glycogenin (GNN) in Neurospora crassa. This work presents a set of biochemical and functional studies confirming the GNN role in glycogen biosynthesis. Kinetic experiments showed a very low GNN K-m (4.41 mu M) for the substrate UDP-glucose. Recombinant GNN was produced in Escherichia coli and analysis by mass spectroscopy identified a peptide containing an oligosaccharide chain attached to Tyr196 residue. Site-directed mutagenesis and functional complementation of a Saccharomyces cerevisiae mutant strain confirmed the participation of this residue in the GNN self-glucosylation and indicated the Tyr198 residue as an additional, although less active, glucosylation site. The physical interaction between GNN and glycogen synthase (GSN) was analyzed by the two-hybrid assay. While the entire GSN was required for full interaction, the C-terminus in GNN was more important. Furthermore, mutation in the GNN glucosylation sites did not impair the interaction with GSN. (c) 2005 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies.en
dc.description.affiliationUNESP, Inst Quim, Dept Bioquim & Tecnol Quim, BR-14800900 Araraquara, SP, Brazil
dc.description.affiliationIndiana Univ, Sch Med, Dept Biochem & Mol Biol, Indianapolis, IN 46202 USA
dc.description.affiliationUSP, Fac Filosofia Ciências & Letras, Dept Biol, BR-14040901 Ribeirao Preto, SP, Brazil
dc.description.affiliationUnespUNESP, Inst Quim, Dept Bioquim & Tecnol Quim, BR-14800900 Araraquara, SP, Brazil
dc.format.extent2208-2214
dc.identifierhttp://dx.doi.org/10.1016/j.febslet.2005.02.075
dc.identifier.citationFebs Letters. Amsterdam: Elsevier B.V., v. 579, n. 10, p. 2208-2214, 2005.
dc.identifier.doi10.1016/j.febslet.2005.02.075
dc.identifier.fileWOS000228310700032.pdf
dc.identifier.issn0014-5793
dc.identifier.lattes8817669953838863
dc.identifier.urihttp://hdl.handle.net/11449/31644
dc.identifier.wosWOS:000228310700032
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofFEBS Letters
dc.relation.ispartofsjr1,991
dc.rights.accessRightsAcesso aberto
dc.sourceWeb of Science
dc.subjectglycogenpt
dc.subjectglycogeninpt
dc.subjectsite-directed mutagenesispt
dc.subjectmass spectrometrypt
dc.subjectbeast complementationpt
dc.subjectNeurospora crassapt
dc.titleBiochemical characterization of Neurospora crassa glycogenin (GNN), the self-glucosylating initiator of glycogen synthesisen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderElsevier B.V.
unesp.author.lattes8817669953838863
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Química, Araraquarapt

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