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α-微管蛋白II是恶性疟原虫中的一种雄性特异性蛋白。

Alpha-tubulin II is a male-specific protein in Plasmodium falciparum.

作者信息

Rawlings D J, Fujioka H, Fried M, Keister D B, Aikawa M, Kaslow D C

机构信息

Laboratory of Malaria Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.

出版信息

Mol Biochem Parasitol. 1992 Dec;56(2):239-50. doi: 10.1016/0166-6851(92)90173-h.

Abstract

The tubulin gene family in Plasmodium falciparum consists of one beta-tubulin and two alpha-tubulin genes (alpha-tubulin I and II). We present here data indicating that alpha-tubulin II is expressed only in male sexual stage parasites. An IgM mAb, 5E7, specifically reacted with stage III (day 4-5) through mature (day 10-11) male gametocytes and with emerging, exflagellating, or freely moving male gametes. No reactivity was detected in female gametocytes, female gametes, sporozoites, or asexual parasites. mAb 5E7 also specifically recognized male gametes of the avian parasite, Plasmodium gallinaceum, and immunoblotted a 50 kDa protein in extracts of male gametes from both species. This 50 kDa antigen was localized by immunoelectron microscopy to axonemes of male gametes in a pattern similar to that obtained with anti-alpha- and anti-beta-tubulin antibodies. Furthermore, mAb 5E7 specifically reacted with recombinant alpha-tubulin II protein obtained using the PCR-amplified alpha-tubulin II gene from a gametocyte-specific cDNA library. The sex-specific expression of alpha-tubulin II and its localization to axoneme of the male parasite suggest a role for this molecule in the morphologic changes that occur during exflagellation and in the motility of the parasite. alpha-Tubulin II and mAb 5E7 may prove useful tools in studies of the biology of sexual stage differentiation and development in P. falciparum in addition to the general understanding of post-translational modifications of tubulin isoforms.

摘要

恶性疟原虫中的微管蛋白基因家族由一个β-微管蛋白基因和两个α-微管蛋白基因(α-微管蛋白I和II)组成。我们在此展示的数据表明,α-微管蛋白II仅在雄性有性阶段寄生虫中表达。一种IgM单克隆抗体5E7,与III期(第4 - 5天)至成熟(第10 - 11天)的雄性配子体以及刚出现、正在进行出丝或自由游动的雄性配子发生特异性反应。在雌性配子体、雌配子、子孢子或无性寄生虫中未检测到反应性。单克隆抗体5E7还特异性识别禽疟原虫(鸡疟原虫)的雄性配子,并在两种物种雄性配子提取物中免疫印迹出一种50 kDa的蛋白质。通过免疫电子显微镜观察,这种50 kDa抗原定位于雄性配子的轴丝上,其模式与使用抗α-和抗β-微管蛋白抗体所获得的模式相似。此外,单克隆抗体5E7与使用来自配子体特异性cDNA文库的PCR扩增α-微管蛋白II基因获得的重组α-微管蛋白II蛋白发生特异性反应。α-微管蛋白II的性别特异性表达及其在雄性寄生虫轴丝上的定位表明,该分子在出丝过程中发生的形态变化以及寄生虫的运动中发挥作用。除了对微管蛋白异构体翻译后修饰的一般理解外,α-微管蛋白II和单克隆抗体5E7可能被证明是研究恶性疟原虫有性阶段分化和发育生物学的有用工具。

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