[Component analysis of excretory/secretory protein from Trichinella spiralis adult worm].

Xiao-di Yang, Zhi-yong Tao, Yang Cheng, Qi Wu, Xiao-li Wang, Di Song, Lan-song Xu, Ren-min Xue, Xue-lian Chang, Hui Zhang, Rui Wang, Xing-zhi Chen, Qiang Fang
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Abstract

Objective: To analyze the component of adult worm excretory/secretory protein(AWESP) from Trichinella spiralis using the shotgun method, and find out the active component underlying its regulatory effect on colitis in humans.

Methods: The T. spiralis AWESP was prepared, separated by SDS-PAGE, lysed with trypsin, and analyzed by shotgun LC-MS/MS. The protein components were determined with the Masco software and classified using the Gene Ontology(GO) method in cellular components, molecular functions, and biological processes.

Results: The AWESPs isolated by SDS-PAGE had a Mr of 15 000-116 000. A total of 280 proteins were revealed by LC-MS/MS, of which 96 were identified by Masco software, 98 were putative, and the remaining 86 were unclear. Preliminary results showed that 4 proteins had regulatory potential for colitis, including cysteine protease inhibitor, serine protease, 53 000 excretory/secretory antigen, and glutathione-S-transferase. GO enrichment analysis showed that the identified proteins had 104 different molecular functions, involved in 363 biological processes.

Conclusion: As revealed by the Masco software, T. spiralis AWESP has complex components and 96 have been identified in this study. Four of them are preliminarily shown to be associated with the anti-colitis effect of T. spiralis.

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旋毛虫成虫排泄/分泌蛋白成分分析
目的:采用散弹法对旋毛虫成虫排泄/分泌蛋白(AWESP)成分进行分析,找出其对人类结肠炎调节作用的有效成分。方法:制备螺旋体AWESP, SDS-PAGE分离,胰蛋白酶裂解,LC-MS/MS分析。使用Masco软件确定蛋白质成分,并使用基因本体(GO)方法对细胞成分、分子功能和生物过程进行分类。结果:SDS-PAGE分离得到的AWESPs的Mr值为15000 ~ 116000。LC-MS/MS共检测到280个蛋白,其中96个为Masco软件鉴定,98个为推测蛋白,86个为未知蛋白。初步结果显示,半胱氨酸蛋白酶抑制剂、丝氨酸蛋白酶、53000排泄/分泌抗原和谷胱甘肽- s转移酶等4种蛋白对结肠炎具有调节作用。氧化石墨烯富集分析表明,鉴定的蛋白具有104种不同的分子功能,参与363个生物过程。结论:经Masco软件分析,螺旋体AWESP成分复杂,共鉴定出96种。其中4种与螺旋锥虫的抗结肠炎作用有关。
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[The effect of Dermatophagoides pteronyssinus group 2 allergen T cell fusion epitope on STAT6 signaling in mice with asthma]. [Component analysis of excretory/secretory protein from Trichinella spiralis adult worm]. [Discovery of Culex inatomii (Diptera : Culicidae) in Chongming, Shanghai]. [The effect of leptin transgenic Plasmodium yoelii on mouse body weight]. [Polymorphism analysis of the block 5 region in merozoite surface protein-1 gene of imported and local Plasmodium vivax in Yunnan Province].
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