期刊
PARASITOLOGY
卷 142, 期 7, 页码 865-878出版社
CAMBRIDGE UNIV PRESS
DOI: 10.1017/S0031182015000190
关键词
microsporidia; Nosema bombycis; surface protein; biotinylation; identification
类别
资金
- National Science and Technology Plan Subject of China (863 Program) [2012AA101301]
- Natural Science Foundation of China [31101770, 31402142]
- Natural Science Foundation Project of Chongqing CSTC [cstc2011jjA80019, cstc2014jcyjA80039]
- Science and Technology Project of the Chongqing Municipal Education Commission of China [KJ100616]
- Natural Science Foundation Project of Chongqing Normal University of China
Parasite-host interactions mediated by cell surface proteins have been implicated as a critical step in infections caused by the microsporidian Nosema bombycis. Such cell surface proteins are considered as promising diagnostic markers and targets for drug development. However, little research has specifically addressed surface proteome identification in microsporidia due to technical barriers. Here, a combined strategy was developed to separate and identify the surface proteins of N. bombycis. Briefly, following (1) biotinylation of the spore surface, (2) extraction of total proteins with an optimized method and (3) streptavidin affinity purification of biotinylated proteins, 22 proteins were identified based on LC-MS/MS analysis. Among them, 5 proteins were confirmed to be localized on the surface of N. bombycis. A total of 8 proteins were identified as hypothetical extracellular proteins, whereas 7 other hypothetical proteins had no available function annotation. Furthermore, a protein with a molecular weight of 18.5kDa was localized on the spore surface by western blotting and immunofluorescence analysis, even though it was predicted to be a nuclear protein by bioinformatics. Collectively, our work provides an effective strategy for isolating microsporidian surface protein components for both drug target identification and further diagnostic research on microsporidian disease control.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据