Title:Proteomic Characterization of the Hyaluronidase (E.C. 3.2.1.35) from the Venom of the Social Wasp Polybia paulista
Volume: 19
Issue: 6
Author(s): Jose Roberto Aparecido dos Santos Pinto, Lucilene Delazari dos Santos, Helen Andrade Arcuri, Nathalia Baptista Dias and Mario Sergio Palma
Affiliation:
Keywords:
Allergen, hyaluronidase, mass spectrometry, molecular modeling, peptide sequencing, wasp venom, Hymenoptera insects, phospholipases, extracellular matrix, post-translational modifications
Abstract: Polybia paulista wasp venom possesses three major allergens: phospholipase A1, 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 (β/α)5 core with alternation
of β-strands and α-helices; the tertiary structure stabilized by a single disulfide bridge between the residues
Cys189 and Cys201. 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”.
