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Development of a strategy for the identification of surface proteins in the pathogenic microsporidian Nosema bombycis

Published online by Cambridge University Press:  26 March 2015

WEIXI ZHAO ,
YOUJIN HAO ,
LINGLIN WANG ,
ZEYANG ZHOU  and
ZHI LI
WEIXI ZHAO
Affiliation:
College of Life Sciences, Chongqing Normal University, Chongqing, China
YOUJIN HAO
Affiliation:
College of Life Sciences, Chongqing Normal University, Chongqing, China
LINGLIN WANG
Affiliation:
College of Life Sciences, Chongqing Normal University, Chongqing, China
ZEYANG ZHOU
Affiliation:
College of Life Sciences, Chongqing Normal University, Chongqing, China The State Key Laboratory of Silkworm Genome Biology, Southwest University, Chongqing, China
ZHI LI*
Affiliation:
College of Life Sciences, Chongqing Normal University, Chongqing, China
*
*Corresponding author. College of Life Sciences, Chongqing Normal University, Chongqing 401331, China. E-mail: [email protected]
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Summary

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 18·5 kDa 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.

Keywords

microsporidiaNosema bombycissurface proteinbiotinylationidentification
Type
Research Article
Information
Parasitology , Volume 142 , Issue 7 , June 2015 , pp. 865 - 878
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Copyright
Copyright © Cambridge University Press 2015 

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