Kinetic and mechanistic characterization of the Sphingomyelinases D from Loxosceles intermedia spider venom

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dc.contributor.authorde Andrade, S. A.
dc.contributor.authorMurakami, M. T.
dc.contributor.authorCavalcante, D. P.
dc.contributor.authorArni, R. K.
dc.contributor.authorTambourgi, D. V.
dc.contributor.institutionInstituto Butantan
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2014-05-20T15:23:10Z
dc.date.available2014-05-20T15:23:10Z
dc.date.issued2006-03-15
dc.description.abstractEnvenomation by arachnids of the genus Loxosceles leads to local dermonecrosis and serious systemic toxicity mainly induced by sphingomyelinases D (SMase D). These enzymes catalyze the hydrolysis of sphingomyelin resulting in the formation of ceramide-phosphate and choline as well as the cleavage of lysophosphatidyl choline generating the lipid mediator lysophosphatidic acid. We have, previously, cloned and expressed two functional SMase D isoforms, named P1 and P2, from Loxosceles intertnedia venom and comparative protein sequence analysis revealed that they are highly homologous to SMase I from Loxosceles laeta which folds to form an (alpha/beta)(8) barrel. In order to further characterize these proteins, pH dependence kinetic experiments and chemical modification of the two active SMases D isoforms were performed. We show here that the amino acids involved in catalysis and in the metal ion binding sites are strictly conserved in the SMase D isoforms from L. intermedia. However, the kinetic studies indicate that SMase P1 hydrolyzes sphingomyelin less efficiently than P2, which can be attributed to a substitution at position 203 (Pro-Leu) and local amino acid substitutions in the hydrophobic channel that could probably play a role in the substrate recognition and binding. (c) 2005 Elsevier Ltd. All rights reserved.en
dc.description.affiliationInst Butantan, Lab Imunoquim, BR-05503900 São Paulo, Brazil
dc.description.affiliationUNESP, IBILCE, Dept Fis, BR-15054000 Sao Jose do Rio Preto, Brazil
dc.description.affiliationUnespUNESP, IBILCE, Dept Fis, BR-15054000 Sao Jose do Rio Preto, Brazil
dc.format.extent380-386
dc.identifierhttp://dx.doi.org/10.1016/j.toxicon.2005.12.005
dc.identifier.citationToxicon. Oxford: Pergamon-Elsevier B.V., v. 47, n. 4, p. 380-386, 2006.
dc.identifier.doi10.1016/j.toxicon.2005.12.005
dc.identifier.issn0041-0101
dc.identifier.lattes9162508978945887
dc.identifier.orcid0000-0003-2460-1145
dc.identifier.urihttp://hdl.handle.net/11449/34008
dc.identifier.wosWOS:000236534700002
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofToxicon
dc.relation.ispartofjcr2.352
dc.relation.ispartofsjr0,692
dc.rights.accessRightsAcesso restrito
dc.sourceWeb of Science
dc.subjectLoxosceles venomspt
dc.subjectSphingomyelinase Dpt
dc.subjectsphingomyelinpt
dc.subjectkinetic parameterspt
dc.subjectStructurept
dc.titleKinetic and mechanistic characterization of the Sphingomyelinases D from Loxosceles intermedia spider venomen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderElsevier B.V.
unesp.author.lattes9162508978945887[4]
unesp.author.orcid0000-0003-2460-1145[4]
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências Letras e Ciências Exatas, São José do Rio Pretopt

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Artigos - Física - IBILCE