Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata.
Isolation of biologically active peptides from the venom of Japanese carpenter bee, Xylocopa appendiculata.
复制标题
DOI：
10.1186/s40409-017-0119-6
复制
发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Shinada T
中科院分区：
文献类型：
--
作者：
Kawakami H;Goto SG;Murata K;Matsuda H;Shigeri Y;Imura T;Inagaki H;Shinada T
质谱引导的毒液肽谱分析是一种强大的工具，以高度敏感的方式探索有毒动物的新物质。在这项研究中，这种肽谱分析方法成功地应用于日本孤独木蜂，Xylocopa appendiculata（膜翅目：蜂总目：蜂科：蚁科：木蜂科：木蜂）的毒液肽的研究。虽然有关于木蜂粗毒液的有趣生物学效应的报道，但其肽的结构和生物学功能尚未得到阐明。采用基质辅助激光解吸/电离飞行时间质谱法对尾尾蛇粗毒液进行了多肽谱分析。用反相高效液相色谱法纯化毒液。纯化的多肽经Edman降解、MS/MS分析和/或分子克隆方法进行多肽测序。通过圆二色性分析、脂质体泄漏试验、抗菌、组胺释放和溶血活性试验进行生物学和功能表征。从尾尾蝮蛇毒液中分离到m/z为16508、1939.3和1900.3的新多肽。具有m/z 16508的肽具有分泌磷脂酶A2 （PLA2）同源性，其特征半胱氨酸残基以及在蜜蜂PLA2s中发现的活性位点残基高度保守。两个m/z 1939.3和m/z 1900.3的新肽分别命名为Xac-1和Xac-2。这些肽被发现是两亲性的，并显示抗菌和溶血活性。其效价与从胡蜂毒液中分离得到的mastoparan几乎相同。本文从尾尾鳗毒液中发现了3个新的生物活性肽，分析了它们的分子功能，并比较了它们的序列同源性，探讨了它们的分子多样性。高灵敏度的质谱分析在本研究中发挥了重要作用。本文的在线版本（doi:10.1186/s40409-017-0119-6）包含补充材料，可供授权用户使用。
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. The online version of this article (doi:10.1186/s40409-017-0119-6) contains supplementary material, which is available to authorized users.