期刊
JOURNAL OF PROTEOME RESEARCH
卷 12, 期 7, 页码 3255-3263出版社
AMER CHEMICAL SOC
DOI: 10.1021/pr400110h
关键词
Trypanosoma cruzi; epimastigotes; subproteome; plasma membrane; cell surface trypsinization; biotinylation
资金
- Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq) [563998/2010-5]
- FAPEG (Fundacao de Amparo a Pesquisa do Estado de Goias)
- CAPES (Programa Nacional de Incentivo a Pesquisa em Parasitologia Basica) [23038.005298/2011-83]
- FINEP (Financiadora de Estudos e Projetos)
Trypanosoma cruzi is a protozoan that causes Chagas' disease, a neglected infectious illness that affects millions of people, mostly in Latin America. Here, the cell surface subproteome of the T. cruzi epimastigote life form was characterized. In order to prepare samples enriched in epimastigote plasma membrane protein, two distinct methodologies were optimized and evaluated. The first methodology was based on cell surface trypsinization (Shave) of intact living cells while the second approach used biotinylation of cell surface proteins followed by streptavidin affinity chromatography isolation of the labeled proteins. Both T. cruzi subproteomes were analyzed by LC-MS/MS. The results showed that the methodologies offered comprehensive and complementary information about the parasite's plasma membrane subproteome.
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