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

dc.contributorUniversidade Estadual Paulista (UNESP)
dc.creatorJusto Jacomini, Débora Laís
dc.creatorCampos Pereira, Franco Dani
dc.creatorAparecido dos Santos Pinto, José Roberto
dc.creatordos Santos, Lucilene Delazari
dc.creatorda Silva Neto, Antonio Joaquim
dc.creatorGiratto, Danielli Thieza
dc.creatorPalma, Mario Sergio
dc.creatorde Lima Zollner, Ricardo
dc.creatorBrochetto Braga, Márcia Regina
dc.date2014-05-27T11:28:40Z
dc.date2016-10-25T18:45:36Z
dc.date2014-05-27T11:28:40Z
dc.date2016-10-25T18:45:36Z
dc.date2013-03-15
dc.date.accessioned2017-04-06T02:16:46Z
dc.date.available2017-04-06T02:16:46Z
dc.identifierToxicon, v. 64, p. 70-80.
dc.identifier0041-0101
dc.identifier1879-3150
dc.identifierhttp://hdl.handle.net/11449/74837
dc.identifierhttp://acervodigital.unesp.br/handle/11449/74837
dc.identifier10.1016/j.toxicon.2012.12.019
dc.identifierWOS:000315706400010
dc.identifier2-s2.0-84873175551.pdf
dc.identifier2-s2.0-84873175551
dc.identifierhttp://dx.doi.org/10.1016/j.toxicon.2012.12.019
dc.identifier.urihttp://repositorioslatinoamericanos.uchile.cl/handle/2250/895596
dc.descriptionIn 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.
dc.languageeng
dc.relationToxicon
dc.rightsinfo:eu-repo/semantics/openAccess
dc.subjectCDNA cloning
dc.subjectHyaluronidase
dc.subjectPolybia paulista venom
dc.subjectPp-Hyal-specific antibody
dc.subjectProtein purification
dc.subjectStructural modeling
dc.subjectcomplementary DNA
dc.subjectglycosidase
dc.subjecthyaluronidase
dc.subjectpolyclonal antibody
dc.subjectwasp venom
dc.subjectAgelaia pallipes pallipes
dc.subjectamino acid composition
dc.subjectApis mellifera
dc.subjectcross reaction
dc.subjectDNA sequence
dc.subjectendemic species
dc.subjectHymenoptera
dc.subjectimmunoblotting
dc.subjectmass spectrometry
dc.subjectmolecular cloning
dc.subjectmolecular weight
dc.subjectnonhuman
dc.subjectPolistes annularis
dc.subjectPolistes lanio lanio
dc.subjectPolybia ignobilis
dc.subjectPolybia paulista
dc.subjectPolybia sericea
dc.subjectpriority journal
dc.subjectprotein glycosylation
dc.subjectprotein purification
dc.subjectprotein structure
dc.subjectproteomics
dc.subjectsequence analysis
dc.subjectsolenopsis invicta
dc.subjectstructure analysis
dc.subjectVespidae
dc.subjectAmino Acid Sequence
dc.subjectAnimals
dc.subjectBase Sequence
dc.subjectBees
dc.subjectCloning, Molecular
dc.subjectCross Reactions
dc.subjectDNA, Complementary
dc.subjectHyaluronoglucosaminidase
dc.subjectMolecular Sequence Data
dc.subjectMolecular Weight
dc.subjectProtein Structure, Tertiary
dc.subjectProteomics
dc.subjectSequence Alignment
dc.subjectSpecies Specificity
dc.subjectWasp Venoms
dc.subjectWasps
dc.titleHyaluronidase from the venom of the social wasp Polybia paulista (Hymenoptera, Vespidae): Cloning, structural modeling, purification, and immunological analysis
dc.typeOtro


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