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

Journal of venomous animals and toxins

Kazuya Murata¹  Tetsuro Shinada²  Hideaki Matsuda³  Yasushi Shigeri³  Hiroko Kawakami⁴  Shin G. Goto⁵  Tomohiro Imura⁶  Hidetoshi Inagaki⁷ 

[1] Geosciences, Osaka City University, Sumiyoshi, Japan;Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki, Japan;Faculty of Pharmacy, Kindai University, Higashiosaka, Japan;Graduate School of Material Science, Osaka City University, Sumiyoshi, Japan;Graduate School of Science, Department of Biology &Health Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Osaka, Japan;Research Institute for Chemical Process Technology, National Institute of Advanced Industrial Science and Technology (AIST), Ibaraki, Japan

关键词: Xylocopa appendiculata;    Carpenter bee;    Venom peptides;    Solitary bee;    Mass spectrometry analysis;   

DOI  :  10.1186/s40409-017-0119-6

学科分类：药理学

来源: BioMed Central

【 摘 要 】

Mass 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. The 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. Three 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. We 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.

【 授权许可】

CC BY