title: 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.
authors: College of Life Sciences,Chongqing Normal University,Chongqing,China.
authors: Weixi
authors: W
authors: Zhao
authors: College of Life Sciences,Chongqing Normal University,Chongqing,China.
authors: Youjin
authors: Y
authors: Hao
authors: College of Life Sciences,Chongqing Normal University,Chongqing,China.
authors: Linglin
authors: L
authors: Wang
authors: College of Life Sciences,Chongqing Normal University,Chongqing,China.
authors: Zeyang
authors: Z
authors: Zhou
authors: College of Life Sciences,Chongqing Normal University,Chongqing,China.
authors: Zhi
authors: Z
authors: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019921:Biotinylation; D012831:Bombyx / Q000382:microbiology; D002846:Chromatography, Affinity; D002853:Chromatography, Liquid; D005656:Fungal Proteins / Q000302:isolation & purification*; D006790:Host-Parasite Interactions; D007814:Larva / Q000382:microbiology; D008565:Membrane Proteins / Q000302:isolation & purification*; D058977:Molecular Sequence Annotation; D016818:Nosema / Q000737:chemistry* / Q000472:pathogenicity / Q000502:physiology; D020543:Proteome / Q000302:isolation & purification*; D013172:Spores, Fungal / Q000737:chemistry* / Q000378:metabolism / Q000472:pathogenicity; D019809:Streptavidin / Q000737:chemistry; D053719:Tandem Mass Spectrometry
chemical_list: D005656:Fungal Proteins; D008565:Membrane Proteins; D020543:Proteome; D019809:Streptavidin
keywords: Nosema bombycis; biotinylation; identification; microsporidia; surface protein