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dc.contributor.authorCano, Veridiana S. P.-
dc.contributor.authorMedrano, Francisco Javier-
dc.contributor.authorPark, Myung Hee-
dc.contributor.authorValentini, Sandro Roberto-
dc.identifier.citationAmino Acids. New York: Springer, v. 38, n. 2, p. 479-490, 2010.-
dc.description.abstractThe unique amino acid hypusine is formed exclusively in eIF5A by the successive action of deoxyhypusine synthase and deoxyhypusine hydroxylase (yeast Lia1, human DOHH). Although the first enzyme has been extensively studied, both Lia1 structure and the mechanism of action remain unclear. Hence, a multi-approach was used to evaluate Lia1 catalysis, metal/substrate binding, structural conformation and stability. Mutational analyses of Lia1 revealed fine differences in the mode of substrate binding between the human and yeast counterparts. Like human DOHH, recombinant Lia1 is an iron metalloenzyme. Iron is essential for enzyme activity since its loss renders the enzyme totally inactive. The separation of iron-free and iron-bound forms by gel filtration and native electrophoresis suggests differences in Lia1 tertiary structure related to the iron binding. The ability of Lia1 to undergo conformational changes prompted us to use a set of complementary spectroscopic approaches and SAXS to obtain detailed information on the processes underlying dissociation of iron from Lia1 at different levels of the protein organization. The additive effect of weak interactions, especially within the metal center, resulted in an active enzyme in a stabilized and compact three-dimensional fold. Loss of tertiary contacts upon iron displacement led to an elongated conformation of Lia1, in which the N- and C-terminal domains are no longer in close proximity to guarantee the proper orientation of the active groups within the active site pocket. Our results demonstrate an essential structural role for iron binding in addition to its contribution to the catalysis of hypusine formation in the eIF-5A precursor.en
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)-
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)-
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)-
dc.sourceWeb of Science-
dc.subjectDeoxyhypusine hydroxylaseen
dc.subjectHEAT-repeat containing proteinen
dc.subjectIron metalloenzymeen
dc.subjectStructural analysisen
dc.titleEvidence for conformational changes in the yeast deoxyhypusine hydroxylase Lia1 upon iron displacement from its active siteen
dc.contributor.institutionUniversidade Estadual Paulista (UNESP)-
dc.contributor.institutionNatl Lab Synchrotron Light-
dc.contributor.institutionNatl Inst Dent & Craniofacial Res-
dc.description.affiliationSão Paulo State Univ UNESP, Dept Biol Sci, Sch Pharmaceut Sci, BR-14801902 Araraquara, SP, Brazil-
dc.description.affiliationNatl Lab Synchrotron Light, BR-13803100 Campinas, SP, Brazil-
dc.description.affiliationNatl Inst Dent & Craniofacial Res, Oral & Pharyngeal Canc Branch, NIH, Bethesda, MD 20892 USA-
dc.description.affiliationUnespSão Paulo State Univ UNESP, Dept Biol Sci, Sch Pharmaceut Sci, BR-14801902 Araraquara, SP, Brazil-
dc.rights.accessRightsAcesso restrito-
dc.relation.ispartofAmino Acids-
Appears in Collections:Artigos, TCCs, Teses e Dissertações da Unesp

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