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

Abstract:

: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.

journal_name

Parasitology

journal_title

Parasitology

authors

Zhao W,Hao Y,Wang L,Zhou Z,Li Z

doi

10.1017/S0031182015000190

subject

Has Abstract

pub_date

2015-06-01 00:00:00

pages

865-78

issue

7

eissn

0031-1820

issn

1469-8161

pii

S0031182015000190

journal_volume

142

pub_type

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