Artigo
Using Proteomic Strategies for Sequencing and Post-Translational Modifications Assignment of Antigen-5, a Major Allergen from the Venom of the Social Wasp Polybia paulista
Registro en:
Journal Of Proteome Research. Washington: Amer Chemical Soc, v. 13, n. 2, p. 855-865, 2014.
1535-3893
10.1021/pr4008927
WOS:000331164100045
2901888624506535
3368404126695911
Autor
Aparecido dos Santos-Pinto, Jose Roberto [UNESP]
Santos, Lucilene Delazari dos [UNESP]
Arcuri, Helen Andrade
Castro, Fabio Morato
Kalil, Jorge Elias
Palma, Mario Sergio [UNESP]
Resumen
Antigen-5 is one of the major allergens identified in wasp venoms, and despite the fact that its biological function is still unknown, many studies have demonstrated its allergenicity. In this study, the biochemical and structural characterization of antigen-5 from the venom of the social wasp Polybia paulista are reported. A gel-based mass spectrometry strategy with CID fragmentation methods and classical protocols of protein chemistry, which included N- and C-terminal sequencing, were used to assign the complete sequence and determine the presence/location of the post-translational modifications (PTMs) of this protein. Six different isoforms of antigen-5 were identified in the crude venom of P. paulista; the most abundant, which corresponds to the intact form of this protein, was recognized by the pool of human specific-IgE. This protein was extensively sequenced through CID mass spectrometry, and a series of PTMs were observed such as hydroxylation, phosphorylation, and glycosylation. Sequence data revealed that this protein has 59.3-93.7% identity with antigen-5 proteins from other known vespid venoms. The molecular model of P. paulista antigen-5 shows that this protein has three alpha-helices, one 3(10), helix, and four beta-sheets covering 28 and 17.9% of the sequence, respectively. The identification and characterization of allergenic compounds is essential for the development of advanced component-resolved allergy diagnostics and treatment. 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) Univ Sao Paulo State UNESP, Ctr Study Social Insects, Inst Biosci Rio Claro, Dept Biol, Rio Claro, SP, Brazil Univ Sao Paulo State UNESP, Ctr Study Venoms & Venomous Anim CEVAP, Botucatu, SP, Brazil INCT iii, Sao Paulo, Brazil Discipline Allergy & Immunol HC Incor FMUSP, Sao Paulo, SP, Brazil Univ Sao Paulo State UNESP, Ctr Study Social Insects, Inst Biosci Rio Claro, Dept Biol, Rio Claro, SP, Brazil Univ Sao Paulo State UNESP, Ctr Study Venoms & Venomous Anim CEVAP, Botucatu, SP, Brazil FAPESP: 11/51684-1