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Mass spectrometric characterization of two novel inflammatory peptides from the venom of the social wasp Polybia paulista
Author(s) -
de Souza Bibiana Monson,
Marques Mauricio Ribeiro,
Tomazela Daniela Maria,
Eberlin Marcos Nogueira,
Mendes Maria Anita,
Palma Mario Sergio
Publication year - 2004
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1452
Subject(s) - chemistry , venom , chromatography , peptide , mass spectrometry , tandem mass spectrometry , isobaric labeling , quadrupole time of flight , protein mass spectrometry , biochemistry
The social wasp P. paulista is relatively common in southeast Brazil causing many medically important stinging incidents. The seriousness of these incidents is dependent on the amount of venom inoculated by the wasps into the victims, and the characteristic envenomation symptoms are strongly dependent on the types of peptides present in the venom. In order to identify some of these naturally occurring peptides available in very low amounts, an analytical protocol was developed that uses a combination of reversed‐phase and normal‐phase high‐performance liquid chromatography (HPLC) of wasp venom for peptide purification, with matrix‐assisted laser desorption/ionization time‐of‐flight post‐source decay mass spectrometry (MALDI‐Tof‐PSD‐MS) and low‐energy collision‐induced dissociation (CID) in a quadrupole time‐of‐flight tandem mass spectrometry (QTof‐MS/MS) instrument for peptide sequencing at the sub‐picomole level. The distinction between Leu and Ile was achieved both by observing d ‐type fragment ions obtained under CID conditions and by comparison of retention times of the natural peptides and their synthetic counterparts (with different combinations of I and/or L at N‐ and C‐terminal positions). To distinguish the isobaric residues K and Q, acetylation of peptides was followed by Q‐Tof‐MS analysis. The primary sequences obtained were INWLKLGKMVIDAL‐NH 2 (MW 1611.98 Da) and IDWLKLGKMVMDVL‐NH 2 (MW 1658.98 Da). Micro‐scale bioassay protocols characterized both peptides as presenting potent hemolytic action, mast cell degranulation, and chemotaxis of polymorphonucleated leukocyte (PMNL) cells. The primary sequences and the bioassay results suggest that these toxins constitute members of a new sub‐class of mastoparan toxins, directly involved in the occurrence of inflammatory processes after wasp stinging. Copyright © 2004 John Wiley & Sons, Ltd.