Laboratory of Molecular Parasitology, Department of Medical Technology, Faculty of Health Sciences, University of Antofagasta, P.O. Box 160, Antofagasta, Chile.
J Biochem Mol Toxicol. 2011 Nov-Dec;25(6):393-403. doi: 10.1002/jbt.20399. Epub 2011 Jun 20.
Toxin phospholipases-D present in the venom of Loxosceles spiders is the principal responsible for local and systemic effects observed in the loxoscelism. In this study, we describe the cloning, expression, functional evaluation, and potential biotechnological application of cDNAs, which code for two new phospholipase D isoforms, LIPLD1 and LIPLD2, of the spider Loxosceles laeta. The recombinant protein rLIPLD1 had hydrolytic activity on sphingomyelin and in vitro hemolytic activity on human red blood cells, whereas rLIPLD2 was inactive. The purified recombinant proteins and the venom are recognized by polyclonal anti-rLIPLD1 and rLIPLD2 sera produced in animals and conferred immunoprotection against the venom. These new isoforms reinforce the importance of the multigene family of phospholipases-D present in Loxosceles spiders. A highly immunogenic inactive isoform such as rLIPLD2 raises important expectation for its use as a potential immunogenic inducer of the immunoprotective response to the toxic action of the venom of Loxosceles laeta.
存在于狼蛛科蜘蛛毒液中的毒素磷脂酶-D 是引起狼蛛咬伤局部和全身效应的主要原因。在这项研究中，我们描述了编码两种新的磷脂酶 D 同工型（LIPLD1 和 LIPLD2）的 cDNA 的克隆、表达、功能评估和潜在的生物技术应用，这些 cDNA 来自于狼蛛 Loxosceles laeta。重组蛋白 rLIPLD1 对神经鞘磷脂具有水解活性，并且在体外对人红细胞具有溶血活性，而 rLIPLD2 则没有活性。纯化的重组蛋白和毒液被在动物中产生的多克隆抗 rLIPLD1 和 rLIPLD2 血清识别，并赋予针对毒液的免疫保护作用。这些新的同工型强化了存在于狼蛛科蜘蛛中的多基因家族磷脂酶-D 的重要性。像 rLIPLD2 这样一种高度免疫原性的无活性同工型，为其作为潜在的免疫原性诱导剂，诱导对 Loxosceles laeta 毒液的毒性作用的免疫保护反应，带来了重要的期望。
