Institute of Biosciences of Rio Claro, Department of Biology, Center of the Study of Social Insects/Dept. Biology, University of São Paulo State (UNESP), Rio Claro, SP, Brazil.
Proteomics. 2011 Apr;11(8):1403-12. doi: 10.1002/pmic.201000414. Epub 2011 Feb 25.
The phospholipases A(1) (PLA(1) s) from the venom of the social wasp Polybia paulista occur as a mixture of different molecular forms. To characterize the molecular origin of these structural differences, an experimental strategy was planned combining the isolation of the pool of PLAs from the wasp venom with proteomic approaches by using 2-D, MALDI-TOF-TOF MS and classical protocols of protein chemistry, which included N- and C-terminal sequencing. The existence of an intact form of PLA(1) and seven truncated forms was identified, apparently originating from controlled proteolysis of the intact protein; in addition to this, four of these truncated forms also presented carbohydrates attached to their molecules. Some of these forms are immunoreactive to specific-IgE, while others are not. These observations permit to raise the hypothesis that naturally occurring proteolysis of PLA(1) , combined with protein glycosylation may create a series of different molecular forms of these proteins, with different levels of allergenicity. Two forms of PLA(2) s, apparently related to each other, were also identified; however, it was not possible to determine the molecular origin of the differences between both forms, except that one of them was glycosylated. None of these forms were immunoreactive to human specific IgE.
来自社交黄蜂 Polybia paulista 的磷脂酶 A(1)(PLA(1) s)以不同分子形式的混合物形式存在。为了表征这些结构差异的分子起源,计划采用分离黄蜂毒液中 PLA 池的实验策略,并结合使用 2-D、MALDI-TOF-TOF MS 和蛋白质化学的经典方法,包括 N-和 C-末端测序。确定存在完整形式的 PLA(1)和七种截断形式,显然源自完整蛋白质的受控蛋白水解;除此之外,其中四种截断形式的分子也连接有碳水化合物。这些形式中的一些对特异性 IgE 具有免疫反应性,而其他形式则没有。这些观察结果使我们提出假设,即 PLA(1) 的天然发生的蛋白水解作用,与蛋白质糖基化相结合,可能会产生这些蛋白质的一系列不同的分子形式,具有不同的致敏性。还鉴定了两种似乎相互关联的 PLA(2) s 形式;然而,除了其中一种形式是糖基化的之外,无法确定两种形式之间差异的分子起源。这些形式均对人特异性 IgE 无免疫反应性。