Development of an approach to analyze the interaction between Nosema bombycis (microsporidia) deproteinated chitin spore coats and spore wall proteins.
Development of an approach to analyze the interaction between Nosema bombycis (microsporidia) deproteinated chitin spore coats and spore wall proteins.
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DOI：
10.1016/j.jip.2013.10.004
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发表时间：
2014
影响因子：
3.4
通讯作者：
Donglin Yang;Xiaoqun Dang;R. Tian;Mengxian Long;Chunfeng Li;Tian Li;Jie Chen;Zhi Li;G. Pan;Zeyang Zhou
中科院分区：
文献类型：
--
作者：
Donglin Yang;Xiaoqun Dang;R. Tian;Mengxian Long;Chunfeng Li;Tian Li;Jie Chen;Zhi Li;G. Pan;Zeyang Zhou
家蚕微孢子虫（Nosema bombycis）是寄生于家蚕的一种专性胞内寄生虫。N.家蚕孢子由电子致密的蛋白质外层和电子透明的几丁质内层组成，孢子壁与质膜相连。本研究以家蚕新孢子虫为材料，用1M NaOH溶液煮沸，获得脱蛋白几丁质孢子衣.透射电子显微镜下，壳质孢子衣呈松散卷曲的环状，折射率强，孢子内未观察到细胞器和细胞核。使用抗SWP 25、26、30和32抗体来检测总可溶性和成熟孢子蛋白中的孢子壁蛋白是否可以与DCSC结合。此外，几丁质结合试验表明，在总的可溶性和成熟的孢子蛋白，SWP26，SWP30和SWP32孢子壁蛋白，结合到脱蛋白几丁质孢子外套，虽然SWP25是不能这种相互作用。此外，DCSCs与碱溶性蛋白孵育后，后者通过处理N.用0.1 M NaOH处理。在这种处理之后，SWP32仍然能够结合DCSC，而SWP26和SWP30不能结合。总的来说，DCSC可用于研究孢子壁蛋白的排列，并且它们揭示了微孢子虫孢子壁是如何自组装的。
Nosema bombycis is an obligate intracellular parasite of the Bombyx mori insect. The spore wall ofN. bombycisis composed of an electron-dense proteinaceous outer layer and an electron-transparent chitinous inner layer, and the spore wall is connected to the plasma membrane. In this study, the deproteinated chitin spore coats (DCSCs) were acquired by boilingN.bombycisin 1 M NaOH. Under a transmission electron microscope, the chitin spore coat resembles a loosely curled ring with strong refractivity; organelles and nuclei were not observed inside the spore. The anti-SWP25, 26, 30 and 32 antibodies were used to detect whether spore wall proteins within the total soluble and mature spore proteins could bind to the DCSCs. Furthermore, a chitin binding assay showed that within the total soluble and mature spore proteins, the SWP26, SWP30 and SWP32 spore wall proteins, bound to the deproteinated chitin spore coats, although SWP25 was incapable of this interaction. Moreover, after the DCSCs were incubated with the alkali-soluble proteins, the latter were obtained by treatingN. bombyciswith 0.1 M NaOH. Following this treatment, SWP32 was still capable of binding the DCSCs, while SWP26 and SWP30 were unable to bind. Collectively, the DCSCs are useful for investigating the arrangement of spore wall proteins, and they shed light on how the microsporidia spore wall is self-assembled.