Peptidomic analysis of the venom of the solitary bee Xylocopa appendiculata circumvolans.
Peptidomic analysis of the venom of the solitary bee Xylocopa appendiculata circumvolans.
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单养蜜蜂的毒液的肽分析二甲叶植物的毒液。
DOI：
10.1186/s40409-017-0130-y
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发表时间：
2017
期刊：
影响因子：
--
通讯作者：
Konno K
中科院分区：
文献类型：
--
作者：
Kazuma K;Ando K;Nihei KI;Wang X;Rangel M;Franzolin MR;Mori-Yasumoto K;Sekita S;Kadowaki M;Satake M;Konno K
在HYPOTERAN昆虫毒液中，来自社会黄蜂和蜜蜂的毒液-如蜜蜂，黄蜂和纸蜂-已经有很好的记录。它们的毒液由许多肽和蛋白质组成，用于保护它们的巢穴和自己免受捕食者的伤害。相比之下，独居黄蜂和蜜蜂的毒液尚未成为进一步研究的对象。在独居蜜蜂的情况下，只有少数毒液中的主要肽成分得到了解决。为此，本研究采用液相色谱-质谱联用技术（LC-MS）对旋旋叶木蜂（Xylocopa apendulata rotumvolans）蜂毒的肽组学成分进行了研究。通过反相HPLC以常规方法分离主要肽组分，并通过Edman降解确定其序列，最后通过固相合成证实。使用合成的样品，生物活性（抗菌活性，肥大细胞贬值，溶血，杀利什曼原虫活性）和人工脂双层的孔形成进行了评价。在线质量指纹图谱显示，粗毒液含有124个组分。MS/MS分析得到75个肽组分的全序列。其中大多数与主要的和新的肽木肽有关。其序列GFVALLKKLPLILKHLH-NH 2具有线性阳离子α-螺旋肽的特征;富含疏水性和碱性氨基酸，无二硫键，因此可以预测其采用两亲性α-螺旋二级结构。在生物学评价中，木脂素表现出广谱抗菌活性、中度肥大细胞脱颗粒和杀利什曼原虫活性，但几乎没有溶血活性。此外，该肽能够将孔并入azolectin的人工脂质双层中，证实了通过在生物膜中形成孔而具有细胞溶解活性的机制。LC-ESI-MS和MS/MS分析表明，该蜂毒主要由小肽组成。主要的肽组分，木糖苷和木糖苷，纯化和其特征在于在一个常规的方式。它们的化学和生物学特性与已知的孤蜂毒肽Melectin和Osmin相似，属于线性阳离子α-螺旋肽。孔形成在人工脂质双层首次证明了一个孤立的蜜蜂肽。
Among the hymenopteran insect venoms, those from social wasps and bees – such as honeybee, hornets and paper wasps – have been well documented. Their venoms are composed of a number of peptides and proteins and used for defending their nests and themselves from predators. In contrast, the venoms of solitary wasps and bees have not been the object of further research. In case of solitary bees, only major peptide components in a few venoms have been addressed. Therefore, the aim of the present study was to explore the peptide component profile of the venom from the solitary bee Xylocopa appendiculata circumvolans by peptidomic analysis with using LC-MS. A reverse-phase HPLC connected to ESI-OrbiTrap MS was used for LC-MS. On-line mass fingerprinting was made from TIC, and data-dependent tandem mass spectrometry gave MSMS spectra. A major peptide component was isolated by reverse-phase HPLC by conventional way, and its sequence was determined by Edman degradation, which was finally corroborated by solid phase synthesis. Using the synthetic specimen, biological activities (antimicrobial activity, mast cell devaluation, hemolysis, leishmanicidal activity) and pore formation in artificial lipid bilayer were evaluated. On-line mass fingerprinting revealed that the crude venom contained 124 components. MS/MS analysis gave 75 full sequences of the peptide components. Most of these are related to the major and novel peptide, xylopin. Its sequence, GFVALLKKLPLILKHLH-NH2, has characteristic features of linear cationic α-helical peptides; rich in hydrophobic and basic amino acids with no disulfide bond, and accordingly, it can be predicted to adopt an amphipathic α-helix secondary structure. In biological evaluation, xylopin exhibited broad-spectrum antimicrobial activity, and moderate mast cell degranulation and leishmanicidal activities, but showed virtually no hemolytic activity. Additionally, the peptide was able to incorporate pores in artificial lipid bilayers of azolectin, confirming the mechanism of the cytolytic activity by pore formation in biological membranes. LC-ESI-MS and MS/MS analysis of the crude venom extract from a solitary bee Xylocopa appendiculata circumvolans revealed that the component profile of this venom mostly consisted of small peptides. The major peptide components, xylopin and xylopinin, were purified and characterized in a conventional manner. Their chemical and biological characteristics, belonging to linear cationic α-helical peptides, are similar to the known solitary bee venom peptides, melectin and osmin. Pore formation in artificial lipid bilayers was demonstrated for the first time with a solitary bee peptide.
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影响因子：
4.4
作者：
ORTEGA, E;SCHNEIDER, H;PECHT, I
通讯作者：
PECHT, I
影响因子：
2.9
作者：
WAKAMATSU, K;OKADA, A;HIGASHIJIMA, T
通讯作者：
HIGASHIJIMA, T
影响因子：
4
作者：
Wimley, William C.
通讯作者：
Wimley, William C.
影响因子：
3.4
作者：
Fertig, N;Blick, RH;Behrends, JC
通讯作者：
Behrends, JC
影响因子：
2
作者：
Takahashi, M;Fuchino, H;Sekita, S
通讯作者：
Sekita, S