摘要
【目的】对意蜂工蜂毒腺蜂毒和电取蜂毒蛋白质组成的特点和差异进行探究,为进一步了解蜂毒的化学组成和合理利用提供理论依据。【方法】采用凝胶电泳和非凝胶技术对直接从毒腺内获取蜂毒(毒腺蜂毒)和电取蜂毒的蛋白质组进行比较,对差异蛋白进行质谱分析、功能鉴定和生物信息学分析。【结果】在毒腺蜂毒中,一维凝胶电泳(1-DE)鉴定19种蛋白,双向凝胶电泳(2-DE)鉴定11种蛋白,非凝胶法鉴定14种蛋白,3种方法在毒腺蜂毒中共鉴定30种非冗余蛋白质,它们是蜂毒毒素成分(50%)和保护腺体细胞免受毒素损伤的抗氧化类蛋白和蛋白成熟加工相关的蛋白折叠、分子转运类等(50%)。在电取蜂毒中,1-DE鉴定12种蛋白,2-DE鉴定3种蛋白,非凝胶法鉴定7种蛋白,3种方法共鉴定14种非冗余蛋白质,它们主要是蜂毒毒素成分(93%)。其中类磷脂酶A2在2种蜂毒中首次鉴定,肽基辅氨酰顺反异构酶在毒腺蜂毒中首次发现。蜂毒明肽前蛋白原、镇静肽在毒腺蜂毒中的含量显著高于电取蜂毒。磷脂酶A-2、毒液二肽基肽酶Ⅳ前体、毒液过敏性酸性磷酸酶和肥大细胞脱粒肽在电取蜂毒中含量较高。【结论】毒腺蜂毒中蛋白种类丰富,但电取蜂毒中主要毒素成分含量不低于毒腺蜂毒。由于蜂毒中的主要药物成分在毒素中,因此,电击取毒可有效利用蜂毒的功能成分。新发现的蜂毒蛋白对进一步认识蜂毒组成成分具有一定意义,这为蜂毒合理利用提供理论依据和实践基础。
[Objective] The objective of this study is to investigate bee venom composition and difference by comparison of bee venom collected from venom glands and electrical stimulated of Italian bees （Apis mellifera ligustica）, and to provide a theoretical basis for pharmaceutical application. [Method] The proteome between bee venom collected directly from the venom glands and artificially electrical stimulation were compared using gel-based （one-dimensional gel electrophoresis, 1-DE, two-dimensional gel electrophoresis, 2-DE） and gel-free proteomics approaches, mass spectrometry, and bioinformatics. [Result] In the gland venom, 19, 11 and 14 proteins were identified by 1-DE, 2-DE and shotgun analysis, respectively, which correspond to 30 nonredundant proteins. They were mainly classified as bee venom toxins （50%）, antioxidant systems, protein folding and molecular transporters （50%）. In construct, in electrical stimulated venom, 12, 3 and 7 proteins were identified, respectively, by 1-DE, 2-DE and shotgun analysis. They were classified into 14 nonredundant proteins, which the major components were venom toxins （93%）. Notably, phospholipase A2-1ike protein was identified for the first time in two forms of bee venom and peptidyl-prolyl cis-trans isomerase was identified only in the gland venom for the first time. The abundance of apamin preproprotein and secapin in the gland venom were significantlyhigher than those in the electrical stimulated venom. However, phospholipase A-2, venom dipeptidyl peptidase IV precursor, venom allergen acid phosphatase and mast cell degranulating peptide had higher abundance in the electrical stimulated venom than those in the gland venom. [ Conclusion ] The venom collected from venom glands contains more protein species, but the abundance of the toxin proteins in electrical stimulated venom are no less than the venom collected from the glands. As the pharmacological components are mainly contained in the toxin proteins, the electrical stimulated venom is a convenient and effective way for utilization. The identified new proteins significantly extend the knowledge of bee venom composition. The result may provide a theoretical and practical basis for future rational use of the honeybee venom.
出处
《中国农业科学》
CAS
CSCD
北大核心
2013年第7期1448-1462,共15页
Scientia Agricultura Sinica
基金
国家现代农业产业技术体系(蜜蜂)(CARS-45-KXJ13)
"十二五"国家科技支撑计划(2011BAD33B04)
关键词
意大利蜜蜂
毒腺蜂毒
电取蜂毒
蛋白质组
凝胶电泳
非凝胶电泳
Apis mellifera ligustica
bee venom directly collected from venom glands
bee venom collected from electricalstimulated
proteome
gel-electrophoresis
gel-free
