Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata

Abstract

BackgroundMass spectrometry-guided venom peptide profiling is a powerful tool to explore novel substances from venomous animals in a highly sensitive manner. In this study, this peptide profiling approach is successfully applied to explore the venom peptides of a Japanese solitary carpenter bee, Xylocopa appendiculata (Hymenoptera: Apoidea: Apidae: Anthophila: Xylocopinae: Xylocopini). Although interesting biological effects of the crude venom of carpenter bees have been reported, the structure and biological function of the venom peptides have not been elucidated yet.MethodsThe venom peptide profiling of the crude venom of X. appendiculata was performed by matrix-assisted laser desorption/ionization-time of flight mass spectroscopy. The venom was purified by a reverse-phase HPLC. The purified peptides were subjected to the Edman degradation, MS/MS analysis, and/or molecular cloning methods for peptide sequencing. Biological and functional characterization was performed by circular dichroism analysis, liposome leakage assay, and antimicrobial, histamine releasing and hemolytic activity tests.ResultsThree novel peptides with m/z 16508, 1939.3, and 1900.3 were isolated from the venom of X. appendiculata. The peptide with m/z 16508 was characterized as a secretory phospholipase A2 (PLA2) homolog in which the characteristic cysteine residues as well as the active site residues found in bee PLA2s are highly conserved. Two novel peptides with m/z 1939.3 and m/z 1900.3 were named as Xac-1 and Xac-2, respectively. These peptides are found to be amphiphilic and displayed antimicrobial and hemolytic activities. The potency was almost the same as that of mastoparan isolated from the wasp venom.ConclusionWe found three novel biologically active peptides in the venom of X. appendiculata and analyzed their molecular functions, and compared their sequential homology to discuss their molecular diversity. Highly sensitive mass analysis plays an important role in this study.Electronic supplementary materialThe online version of this article (doi:10.1186/s40409-017-0119-6) contains supplementary material, which is available to authorized users.

Evaluation of the bioactivity of a mastoparan peptide from wasp venom and of its analogues designed through targeted engineering

“…Pharmacologically, mastoparans also represent a promising group of small peptides, with those mastoparans that have been characterised variously displaying broad-spectrum action against microorganisms, inhibitory effects against tumour proliferation, and stimulating serotonin release from platelets and mast cell degranulation 10 - 12 . Effective active concentrations are also often low, particularly for antimicrobial activity where suitable inhibitory concentrations are on the order of micromolars or even lower 13 - 15 .…”

Dipteran endoparasitoid Exorista bombycis utilizes antihemocyte components against host defense of silkworm Bombyx mori

“…Enhanced expression of HAIP gene in the parasitoid integument, subsequent presence of HAIP in E. bombycis tissue protein, In addition, peptides of a few proteins associated with cellular ill-functions are also identified from E. bombycis larval tissue (Table 2). The braconid Aphidius ervi produced venom rich in γ-glutamyl transpeptidases (γ-GTs) and cysteine-rich toxin-like peptides (Colinet et al, 2014), Xylocopa appendiculata produced secretory Phospholipase A2 homolog, Xac-1 and Xac-2 (Kawakami et al, 2017) and Asobara tabida produced hydrolases with sequence similarity to aspartyl glucosaminidase subunits (Moreau et al, 2004). VPr3, a 312 amino acid peptide component of toxin protein with antihemocyte aggregation properties, was identified from P. hypochondriaca (Richards & Dani, 2008).…”

A common venomous ancestor? Prevalent bee venom genes evolved before the aculeate stinger while few major toxins are bee-specific

“…Melittin-like sequences in other Apis species (A. dorsata, A. cerana and A. florea) feature a similar microsyntenic arrangement (Figure 4). Other bee species also possess melittin-like sequences (bombolittin, osmin, collectin, lasioglossin, melectin and macropin (Cerovský et al 2008;Cerovský et al 2009;Stöcklin et al 2010;Čujová et al 2014;Monincová et al 2014;Kawakami et al 2017). Microsynteny analysis provided evidence that they are orthologous in at least some species from the genera Colletes, Osmia and Bombus (Figure 4).…”