Proteomic Analysis of Trypanosoma cruzi Secretome: Characterization of Two Populations of Extracellular Vesicles and Soluble Proteins

Proteomic Analysis of Trypanosoma cruzi Secretome: Characterization of Two Populations of Extracellular Vesicles and Soluble Proteins

Author Bayer-Santos, Ethel Autor UNIFESP Google Scholar
Aguilar-Bonavides, Clemente Google Scholar
Rodrigues, Silas Pessini Google Scholar
Cordero, Esteban Mauricio Google Scholar
Marques, Alexandre Ferreira Google Scholar
Varela-Ramirez, Armando Google Scholar
Choi, Hyungwon Google Scholar
Yoshida, Nobuko Autor UNIFESP Google Scholar
Silveira, Jose Franco da Autor UNIFESP Google Scholar
Almeida, Igor C. Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Texas El Paso
Natl Univ Singapore
Abstract Microorganisms use specialized systems to export virulence factors into host cells. Secretion of effector proteins into the extracellular environment has been described in Trypanosoma cruzi; however, a comprehensive proteomic analysis of the secretome and the secretion mechanisms involved remain elusive. Here, we present evidence that T. cruzi releases proteins associated with vesicles that are formed by at least two different mechanisms. Transmission electron microscopy showed larger vesicles budding from the plasma membrane of noninfective epimastigotes and infective meta-cyclic trypomastigotes, as well as smaller vesicles within the flagellar pocket of both forms. Parasite conditioned culture supernatant was fractionated and characterized by morphological, immunochemical, and proteomic analyses. Three fractions were obtained by differential ultracentrifugation: the first enriched in larger vesicles resembling ectosomes, the second enriched in smaller vesicles resembling exosomes, and a third fraction enriched in soluble proteins not associated with extracellular vesicles. Label-free quantitative proteomic analysis revealed a rich collection of proteins involved in metabolism, signaling, nucleic acid binding, and parasite survival and virulence. These findings support the notion that T. cruzi uses different secretion pathways to excrete/secrete proteins. Moreover, our results suggest that metacyclic forms may use extracellular vesicles to deliver cargo into host cells.
Keywords Trypanosoma cruzi
protozoan parasite
Chagas disease
secretion pathways
secretome
extracellular vesicles
exosomes
ectosomes
proteomic analysis
Language English
Sponsor National Institutes of Health (NIH)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Research Centers in Minority Institutions (RCMI) program
National Institutes on Minority Health and Health Disparities (NIMHD), a component of the NIH
Grant number National Institutes of Health (NIH): R01AI070655-A5
National Institutes of Health (NIH): R01AI070655-A5S1
Research Centers in Minority Institutions (RCMI) program: 8G12MD007592
Research Centers in Minority Institutions (RCMI) program: 5G12RR008124-16A1
Research Centers in Minority Institutions (RCMI) program: 5G12RR008124-16A1S1
Date 2013-02-01
Published in Journal of Proteome Research. Washington: Amer Chemical Soc, v. 12, n. 2, p. 883-897, 2013.
ISSN 1535-3893 (Sherpa/Romeo, impact factor)
Publisher Amer Chemical Soc
Extent 883-897
Origin http://dx.doi.org/10.1021/pr300947g
Access rights Closed access
Type Article
Web of Science ID WOS:000314558800030
URI http://repositorio.unifesp.br/handle/11600/35951

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