Two new phospholipase D isoforms of Loxosceles laeta: cloning, heterologous expression, functional characterization, and potential biotechnological application.

Catalan A; Cortes W; Sagua H; Gonzalez J; Araya JE

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Two new phospholipase D isoforms of Loxosceles laeta: cloning, heterologous expression, functional characterization, and potential biotechnological application.

机译：乳突线虫的两种新的磷脂酶D同工型：克隆，异源表达，功能表征和潜在的生物技术应用。

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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.

机译：存在于蛇颈蜘蛛毒液中的毒素磷脂酶-D是导致在蛇颈龙中观察到的局部和全身作用的主要原因。在这项研究中，我们描述了cDNA的克隆，表达，功能评估和潜在的生物技术应用，这些cDNA编码了蜘蛛夜蛾的两个新的磷脂酶D亚型LIPLD1和LIPLD2。重组蛋白rLIPLD1对鞘磷脂具有水解活性，对人红细胞具有体外溶血活性，而rLIPLD2没有活性。纯化的重组蛋白和毒液被动物产生的多克隆抗rLIPLD1和rLIPLD2血清识别，并赋予了针对毒液的免疫保护。这些新的同工型加强了在腰椎蜘蛛中存在的磷脂酶-D多基因家族的重要性。高度免疫原性的非活性同工型，例如rLIPLD2，对于其用作对黑尾蛇毒毒性作用的免疫保护反应的潜在免疫原性诱导剂，提出了重要的期望。

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《Journal of biochemical and molecular toxicology》 |2011年第6期|共11页
作者
Catalan A; Cortes W; Sagua H; Gonzalez J; Araya JE;
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正文语种 eng
中图分类生物化学;
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2. Two new phospholipase D isoforms of Loxosceles laeta: cloning, heterologous expression, functional characterization, and potential biotechnological application. [J] . Catalan A, Cortes W, Sagua H, Journal of biochemical and molecular toxicology . 2011,第6期

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Two new phospholipase D isoforms of Loxosceles laeta: cloning, heterologous expression, functional characterization, and potential biotechnological application.

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