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Extending the honey bee venome with the antimicrobial peptide apidaecin and a protein resembling wasp antigen 5

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  • Funding: The authors gratefully acknowledge the Research Foundation of Flanders (FWO-Vlaanderen G041708N and GO62811N) and the University of Leuven Research Foundation (GOA 2010/14) for financial support. M.V.V. and D.C. are funded by the Institute for the Promotion of Innovation through Science and Technology in Flanders (IWT-Vlaanderen). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Correspondence: Matthias Van Vaerenbergh, Laboratory of Zoophysiology, Ghent University, K.L. Ledeganckstraat 35, 9000 Ghent, Belgium. Tel.: +32 9264 5151; fax: +32 9264 5242; e-mail: matthias.vanvaerenbergh@ugent.be

Abstract

Honey bee venom is a complex mixture of toxic proteins and peptides. In the present study we tried to extend our knowledge of the venom composition using two different approaches. First, worker venom was analysed by liquid chromatography-mass spectrometry and this revealed the antimicrobial peptide apidaecin for the first time in such samples. Its expression in the venom gland was confirmed by reverse transcription PCR and by a peptidomic analysis of the venom apparatus tissue. Second, genome mining revealed a list of proteins with resemblance to known insect allergens or venom toxins, one of which showed homology to proteins of the antigen 5 (Ag5)/Sol i 3 cluster. It was demonstrated that the honey bee Ag5-like gene is expressed by venom gland tissue of winter bees but not of summer bees. Besides this seasonal variation, it shows an interesting spatial expression pattern with additional production in the hypopharyngeal glands, the brains and the midgut. Finally, our immunoblot study revealed that both synthetic apidaecin and the Ag5-like recombinant from bacteria evoke no humoral activity in beekeepers. Also, no IgG4-based cross-reactivity was detected between the honey bee Ag5-like protein and its yellow jacket paralogue Ves v 5.

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