Proteomic Characterization of the Hyaluronidase (EC 3.2.1.35) from the Venom of the Social Wasp Polybia paulista


Autoria(s): Aparecido dos Santos Pinto, Jose Roberto; dos Santos, Lucilene Delazari; Arcuri, Helen Andrade; Dias, Nathalia Baptista; Palma, Mario Sergio
Contribuinte(s)

Universidade Estadual Paulista (UNESP)

Data(s)

20/05/2014

20/05/2014

01/06/2012

Resumo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Processo FAPESP: 11/51684-1

Polybia paulista wasp venom possesses three major allergens: phospholipase A(1), hyaluronidase and antigen-5. To the best of our knowledge, no hyaluronidase from the venom of Neotropical social wasps was structurally characterized up to this moment, mainly due to its reduced amount in the venom of the tropical wasp species (about 0.5% of crude venom). Four different glycoproteic forms of this enzyme were detected in the venom of the wasp Polybia paulista. In the present investigation, an innovative experimental approach was developed combining 2-D SDS-PAGE with in-gel protein digestion by different proteolytic enzymes, followed by mass spectrometry analysis under collision-induced dissociation CID) conditions for the complete assignment of the protein sequencing. Thus, the most abundant form of this enzyme in P. paulista venom, the hyaluronidase-III, was sequenced, revealing that the first 47 amino acid residues from the N-terminal region, common to other Hymenoptera venom hyaluronidases, are missing. The molecular modeling revealed that hyaluronidase-III has a single polypeptide chain, folded into a tertiary structure, presenting a central (beta/alpha)(5) core with alternation of beta-strands and alpha-helices; the tertiary structure stabilized by a single disulfide bridge between the residues Cys(189) and Cys(201). The structural pattern reported for P. paulista venom hyaluronidase-III is compatible with the classification of the enzyme as member of the family 56 of glycosidase hydrolases. Moreover, its structural characterization will encourage the use of this protein as a model for future development of component-resolved diagnosis.

Formato

625-635

Identificador

http://eurekaselect.com/97510/article

Protein and Peptide Letters. Sharjah: Bentham Science Publ Ltd, v. 19, n. 6, p. 625-635, 2012.

0929-8665

http://hdl.handle.net/11449/19720

WOS:000304442400008

Idioma(s)

eng

Publicador

Bentham Science Publ Ltd

Relação

Protein and Peptide Letters

Direitos

closedAccess

Palavras-Chave #Allergen #hyaluronidase #mass spectrometry #molecular modeling #peptide sequencing #wasp venom
Tipo

info:eu-repo/semantics/article