鸽毛滴虫外泌体的分离鉴定及蛋白质谱分析

doi:10.11843/j.issn.0366-6964.2020.07.026

Abstract

Abstract: The aim of this study was to determine whether Trichomonas gallinae (T. gallinae) secrete exosomes and investigate the protein composition of exosomes derived from T. gallinae. T. gallinae was isolated from upper digestive tract of infected pigeons and identified by microscopy and sequencing. Exosomes were extracted from serum absent culture medium, and further verified by transmission electron microscopy, Western blot and nanoparticle tracking analysis (NTA). Label-free was used to identify the proteins of exosomes. Results showed that the parasite had T. gallinae morphology, and sequence alignment of ITS1/5.8S/ITS2 gene showed an identity of 98%. The isolated vesicles presented a typical cup-shaped morphology; NTA results showed that particle size concentrated on 125.1 nm, percentage of peak diameter at 132.3 nm was 99.3%; proteins such as CD63 and TSG101 were conspicuously detected, suggesting that the extracts were exosomes. Mass spectrometry results showed that enolase, glyceraldehyde-3-phosphate dehydrogenase, phosphoglycerate mutase and elongation factor were the highly expressed proteins. GO pathways showed that proteins of exosomes mainly enriched into cellular components such as intracellular and cytoplasmic, played GTP binding and GTPase activity function, and participated in biological process of small GTPase mediated signal transduction and glycolytic process. KEGG enrichment analysis showed that proteins of exosomes were enriched in pathways as metabolic pathways, glycolysis/gluconeogenesis, biosynthesis of antibiotics and secondary metabolites. These results indicated that T. gallinae can secrete exosomes and proteins of exosomes might play roles in energy metabolism, signal transduction and biosynthesis.

Key words: pigeon, Trichomonas gallinae, exosomes, Label-free technology, protein profiles

CLC Number:

NI Aixin, LI Yunlei, GE Pingzhuang, WANG Panlin, Adamu Mani Isa, SHI Lei, FAN Jing, SUN Yanyan, SUN Hong, MA Hui, CHEN Jilan. Extraction and Protein Profiling of Exosomes Derived from Trichomonas gallinae in Pigeon[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(7): 1737-1747.

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Extraction and Protein Profiling of Exosomes Derived from Trichomonas gallinae in Pigeon

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