Agelotoxin:: a phospholipase A2 from the venom of the neotropical social wasp cassununga (Agelaia pallipes pallipes) (Hymenoptera-Vespidae)
Agelotoxin:: a phospholipase A2 from the venom of the neotropical social wasp cassununga (Agelaia pallipes pallipes) (Hymenoptera-Vespidae)
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DOI：
10.1016/s0041-0101(99)00199-3
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发表时间：
2000-10-01
期刊：
影响因子：
2.8
通讯作者：
Palma, MS
中科院分区：
文献类型：
--
作者：
Costa, H;Palma, MS
新热带黄蜂Agelaia pallipes pallipes在巴西东南部具有侵略性和地方性，经常在农村地区造成刺痛事故。通过Sephadex G-100凝胶过滤，然后在乙腈/水梯度下在C-18柱中进行高效反相色谱，纯化了agelotoxin：具有磷脂酶A（2）（PLA（2））活性的毒素，其在三种不同聚集状态下发生平衡：单体（mol. wt 14 kDa）、三聚体（mol.重量42 kDa）和五聚体（摩尔。在从单体到五聚体的聚集下，该酶表现出连接到蛋白质链的高糖含量（22% [w/w]）和底物水解的最佳pH值从7.5到9.0的转变。所有的聚集态均呈现Michaelian稳态动力学行为，单体聚合导致磷脂酶活性降低，这是由于四级结构的形成促进了非竞争性抑制。Agelotoxin的PLA（2）催化活性根据其聚集状态而变化（从833 μ mol mg（-1）min（-1）到12533 μ mol mg（-1）min（-1）），单体和寡聚体形式的活性都比来自意大利蜜蜂毒液的PLA（2）和来自胡蜂的角蛋白低。Agelotoxin也是一种非常有效的直接溶血素，其单体的溶血作用比P.保利斯塔的PbTx高200倍，比A.意大利蜜蜂的中性聚乳酸（2）的活性比意大利蜜蜂的中性聚乳酸（2）高570倍。nigricolis，约为眼镜蛇毒心脏毒素的1250倍。(C)2000爱思唯尔科技有限公司版权所有。
The neotropical wasp Agelaia pallipes pallipes is aggressive and endemic in southeast of Brazil, where very often it causes stinging accidents in rural areas. By using gel filtration on Sephadex G-100, followed by high performance reversed phase chromatography in a C-18 column under acetonitrile/water gradient, the agelotoxin was purified: a toxin presenting phospholipase A(2) (PLA(2)) activity, which occurs under equilibrium of three different aggregation states: monomer (mol. wt 14 kDa), trimer (mol. wt 42 kDa) and pentamer (mol. wt 74 kDa).The enzyme presents high sugar contents attached to the protein chain (22% [w/w]) and a transition of the values of pH optimum for the substrate hydrolysis from 7.5 to 9.0, under aggregation from monomer to pentamer. All the aggregation states present Michaelian steady-state kinetic behavior and the monomer polymerization caused a decreasing of phospholipasic activity due a non-competitive inhibition promoted by the formation of a quaternary structure. The PLA(2) catalytic activity of agelotoxin changes according to its state of aggregation (from 833 to 12533 mu mol mg(-1) min(-1)) and both the monomeric and oligomeric forms present lowest activities than the PLA(2) from Apis mellifera venom and hornetin from Vespa basalis. Agelotoxin is also a very potent direct hemolysin; the monomer of agelotoxin presented hemolytic actions until 200 times higher than the PbTx from P. paulista, 740 times higher than the PLA(2) from A. mellifera, 570 times higher than that of neutral PLA(2) from N. nigricolis and about 1250 times than that of cardiotoxin from Naja naja atra venom. (C) 2000 Elsevier Science Ltd. All rights reserved.