Oct. 22-25, 2012
Zhenjiang, China
Proteome Comparison of Honeybee (Apis mellifera ligustica) Worker
Venom Between Directly Collected from Venom Glands and Electrical Stimulations
LI Rong-li, HAN Bin, FANG Yu, FENG Mao, LI Jian-ke
(Institute of Apicultural Research, Chinese Academy of
Agricultural Sciences, Beijing 100093,China) Italian bee (Apis mellifera
ligustica) is the most important bees for the beekeeping industry of China. Understanding
its venom composition is vital for the reasonable use of bee venom for the
human health promotion.
The proteome between the venom collected directly from the
venom glands and artificially electrical stimulations were compared using
gel-based (one-dimensional gel electrophoresis, 1-DE, two-dimensional gel
electrophoresis, 2-DE) and gel-free proteomics approaches, mass spectrometry,
bioinformatics. Overall, 31 nonredundant proteins were identified in the venom
collected from the glands, which are mainly bee venom toxins, antioxidant
system, protein folding, molecular transporters. In construct, only 15 nonredundant
proteins were identified in the venom collected by electrical stimulation,
which mainly contains bee venom toxins. Phospholipase A2-like protein was identified
first time in two forms of venom and peptidyl-prolyl cis-trans isomerase was
identified for the first time only in the venom directly collected from the
glands. There were 17 differentially expressed proteins in two forms of venom
with higher abundance in the venom collected from the gland were protein
folding, molecular transporters, antioxidant systems and part of the venom
toxins, such as melittin, apamin preproprotein, secapin, venom proteases and
icarapin-like precursor.
However, phospholipase A2, allergen Api m 6 and mast cell degranulating
peptide had higher abundance in the venom collected by electrical stimulations.
Our data suggests that the bee venom directly collected from venom glands
potentially has better biological activity and lower allergic reaction than the
electrical stimulations. The identified new proteins significantly increase the
proteome coverage of the bee venom. This may provide theoretical basis for the
rational use of the honeybee venom.
Key words: honeybee; venom; proteome; gel-electrophoresis;
gel-free
意大利工蜂毒腺内蜂毒与电取蜂毒蛋白质组比较
李荣丽，韩 宾，房
宇，冯 毛，李建科
（中国农业科学院蜜蜂研究所，北京 100093，中国）
【目的】意大利蜜蜂（意蜂）是我国饲养主要蜂种，探究其蜂毒组成成分将对合理利用具有重要意义。
【方法】采用凝胶电泳和非凝胶技术对直接从毒腺内取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20种蛋白，双向凝胶电泳（2-DE）鉴定11 种蛋白，非凝胶法鉴定15 种蛋白，三种方法在毒腺蜂毒中共鉴定31 种非冗余蛋白质，它们是蜂毒毒素成分、抗氧化、蛋白折叠、分子转运类等。在电取蜂毒中，1-DE
鉴定13 种蛋白，2-DE
鉴定4 种蛋白，非凝胶方法鉴定8 种蛋白，三种方法共鉴定15 种非冗余蛋白质，主要是蜂毒毒素成分。其中类磷脂酶A2 是在2 种蜂毒中首次鉴定，肽基辅氨酰顺反异构酶是首次在毒腺蜂毒中发现。毒腺蜂毒与电取蜂毒中差异蛋白为17种，其中毒腺蜂毒含量显著高于电取蜂毒的蛋白为蛋白折叠、分子转运类、抗氧化类以及蜂毒毒素成分中的蜂毒肽、蜂毒明肽、镇静肽、毒液蛋白酶、毒液丝氨酸蛋白酶和icarapin-like
precursor。电取蜂毒中含量高于毒腺蜂毒的蛋白为蜂毒毒素成分中的磷脂酶A2、allergen
Api m
6和肥大细胞脱粒肽。【结论】毒腺蜂毒在功能成分的含量上要显著高于电取蜂毒，且过敏源物质低于电取蜂毒，说明毒腺内蜂毒比电取蜂毒不但具有更好的生物学活性而且具有更低的过敏反应，新发现的蛋白是对蜂毒功能成分完善和补充，这将对蜂毒的合理利用提供一定理论依据。
关键词：意大利蜜蜂；毒腺蜂毒；电取蜂毒；蛋白质组；非凝胶电泳；凝胶电泳
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