Hyaluronidase from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae): Cloning, structural modeling, purification, and immunological analysis

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dc.contributor.authorJusto Jacomini, Débora Laís [UNESP]
dc.contributor.authorCampos Pereira, Franco Dani [UNESP]
dc.contributor.authorAparecido dos Santos Pinto, José Roberto [UNESP]
dc.contributor.authordos Santos, Lucilene Delazari
dc.contributor.authorda Silva Neto, Antonio Joaquim
dc.contributor.authorGiratto, Danielli Thieza [UNESP]
dc.contributor.authorPalma, Mario Sergio [UNESP]
dc.contributor.authorde Lima Zollner, Ricardo
dc.contributor.authorBrochetto Braga, Márcia Regina [UNESP]
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.contributor.institutionFazenda Experimental Lageado
dc.contributor.institutionFaculdades Integradas Claretianas
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.date.accessioned2014-05-27T11:28:40Z
dc.date.available2014-05-27T11:28:40Z
dc.date.issued2013-03-15
dc.description.abstractIn this study, we describe the cDNA cloning, sequencing, and 3-D structure of the allergen hyaluronidase from Polybia paulista venom (Pp-Hyal). Using a proteomic approach, the native form of Pp-Hyal was purified to homogeneity and used to produce a Pp-specific polyclonal antibody. The results revealed that Pp-Hyal can be classified as a glycosyl hydrolase and that the full-length Pp-Hyal cDNA (1315 bp; GI: 302201582) is similar (80-90%) to hyaluronidase from the venoms of endemic Northern wasp species. The isolated mature protein is comprised of 338 amino acids, with a theoretical pI of 8.77 and a molecular mass of 39,648.8 Da versus a pI of 8.13 and 43,277.0 Da indicated by MS. The Pp-Hyal 3D-structural model revealed a central core (α/β)7 barrel, two sulfide bonds (Cys 19-308 and Cys 185-197), and three putative glycosylation sites (Asn79, Asn187, and Asn325), two of which are also found in the rVes v 2 protein. Based on the model, residues Ser299, Asp107, and Glu109 interact with the substrate and potential epitopes (five conformational and seven linear) located at surface-exposed regions of the structure. Purified native Pp-Hyal showed high similarity (97%) with hyaluronidase from Polistes annularis venom (Q9U6V9). Immunoblotting analysis confirmed the specificity of the Pp-Hyal-specific antibody as it recognized the Pp-Hyal protein in both the purified fraction and P. paulista crude venom. No reaction was observed with the venoms of Apis mellifera, Solenopsis invicta, Agelaia pallipes pallipes, and Polistes lanio lanio, with the exception of immune cross-reactivity with venoms of the genus Polybia (sericea and ignobilis). Our results demonstrate cross-reactivity only between wasp venoms from the genus Polybia. The absence of cross-reactivity between the venoms of wasps and bees observed here is important because it allows identification of the insect responsible for sensitization, or at least of the phylogenetically closest insect, in order to facilitate effective immunotherapy in allergic patients. © 2013 Elsevier Ltd.en
dc.description.affiliationLab. de Biologia Molecular de Artrópodes-LBMA IBRC-UNESP Univ Estadual Paulista, Av. 24-A, no 1515, CEP 13506-900, Bela Vista, Rio Claro, SP
dc.description.affiliationCentro de Estudos de Insetos Sociais CEIS-IBRC UNESP (Univ Estadual Paulista), Av. 24-A, no 1515, CEP 13506-900, Bela Vista, Rio Claro, SP
dc.description.affiliationFazenda Experimental Lageado, Rua José Barbosa de Barros No. 1780, CEP 18610-307, Botucatu, SP
dc.description.affiliationFaculdades Integradas Claretianas, Av. Sto. Antonio Maria Claret, no 1724, CEP 13503-257, Cidade Claret, Rio Claro, SP
dc.description.affiliationLaboratório de Imunologia and Alergia Experimental-LIAE Faculdade de Ciências Médicas, FCM Universidade Estadual de Campinas-UNICAMP, Cidade Universitaria Zeferino Vaz, Rua Tessalia Vieira de Camargo, no 126, CEP 13083-887, Campinas, SP
dc.description.affiliationUnespLab. de Biologia Molecular de Artrópodes-LBMA IBRC-UNESP Univ Estadual Paulista, Av. 24-A, no 1515, CEP 13506-900, Bela Vista, Rio Claro, SP
dc.description.affiliationUnespCentro de Estudos de Insetos Sociais CEIS-IBRC UNESP (Univ Estadual Paulista), Av. 24-A, no 1515, CEP 13506-900, Bela Vista, Rio Claro, SP
dc.format.extent70-80
dc.identifierhttp://dx.doi.org/10.1016/j.toxicon.2012.12.019
dc.identifier.citationToxicon, v. 64, p. 70-80.
dc.identifier.doi10.1016/j.toxicon.2012.12.019
dc.identifier.file2-s2.0-84873175551.pdf
dc.identifier.issn0041-0101
dc.identifier.issn1879-3150
dc.identifier.lattes2901888624506535
dc.identifier.lattes0316857632230804
dc.identifier.scopus2-s2.0-84873175551
dc.identifier.urihttp://hdl.handle.net/11449/74837
dc.identifier.wosWOS:000315706400010
dc.language.isoeng
dc.relation.ispartofToxicon
dc.relation.ispartofjcr2.352
dc.relation.ispartofsjr0,692
dc.rights.accessRightsAcesso aberto
dc.sourceScopus
dc.subjectCDNA cloningen
dc.subjectHyaluronidaseen
dc.subjectPolybia paulista venomen
dc.subjectPp-Hyal-specific antibodyen
dc.subjectProtein purificationen
dc.subjectStructural modelingen
dc.subjectcomplementary DNAen
dc.subjectglycosidaseen
dc.subjecthyaluronidaseen
dc.subjectpolyclonal antibodyen
dc.subjectwasp venomen
dc.subjectAgelaia pallipes pallipesen
dc.subjectamino acid compositionen
dc.subjectApis melliferaen
dc.subjectcross reactionen
dc.subjectDNA sequenceen
dc.subjectendemic speciesen
dc.subjectHymenopteraen
dc.subjectimmunoblottingen
dc.subjectmass spectrometryen
dc.subjectmolecular cloningen
dc.subjectmolecular weighten
dc.subjectnonhumanen
dc.subjectPolistes annularisen
dc.subjectPolistes lanio lanioen
dc.subjectPolybia ignobilisen
dc.subjectPolybia paulistaen
dc.subjectPolybia sericeaen
dc.subjectpriority journalen
dc.subjectprotein glycosylationen
dc.subjectprotein purificationen
dc.subjectprotein structureen
dc.subjectproteomicsen
dc.subjectsequence analysisen
dc.subjectsolenopsis invictaen
dc.subjectstructure analysisen
dc.subjectVespidaeen
dc.subjectAmino Acid Sequenceen
dc.subjectAnimalsen
dc.subjectBase Sequenceen
dc.subjectBeesen
dc.subjectCloning, Molecularen
dc.subjectCross Reactionsen
dc.subjectDNA, Complementaryen
dc.subjectHyaluronoglucosaminidaseen
dc.subjectMolecular Sequence Dataen
dc.subjectMolecular Weighten
dc.subjectProtein Structure, Tertiaryen
dc.subjectProteomicsen
dc.subjectSequence Alignmenten
dc.subjectSpecies Specificityen
dc.subjectWasp Venomsen
dc.subjectWaspsen
dc.titleHyaluronidase from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae): Cloning, structural modeling, purification, and immunological analysisen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
unesp.author.lattes2901888624506535
unesp.author.lattes0316857632230804
unesp.campusUniversidade Estadual Paulista (Unesp), Instituto de Biociências, Rio Claropt

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Artigos - Biologia - IBRC
Rio Claro - CEIS - Centro de Estudos de Insetos Sociais