Oct. 22-25, 2012
Zhenjiang, ChinaProteome 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
李荣丽，韩 宾，房 宇，冯 毛，李建科
【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