Proteome analysis of the surface of Trichomonas vaginalis reveals novel proteins and strain-dependent differential expression

NATALIA DE MIGUEL; GIL LUSTIG; OLIVIA TWU; ARNAB CHATTOPADHYAY; JAMES A. WOHLSCHLEGEL ; PATRICIA J. JOHNSON

MOLECULAR & CELLULAR PROTEOMICS

AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Año: 2010 vol. 9 p. 1554 - 1566

1535-9476

The identification of surface proteins on the plasma membrane of pathogens is of fundamental importance in understanding host-pathogen interactions. Surface proteins of the extracellular parasite Trichomonas are implicated in the initial adherence to mucosal tissue and are likely toplay a critical role in the long term survival of this pathogen in the urogenital tract. In this study, we used cell surface biotinylation and multidimensional protein identification technology to identify the surface proteome of six strains of Trichomonas vaginalis with differing adherence capacities to vaginal epithelial cells. A combined total of 411 proteins were identified, and of these, 11 were found to be more abundant in adherent strains relative to less adherent parasites. The mRNA levels of five differentially expressed proteins selected for quantitative RT-PCR analysis mirrored their observed protein levels, confirming their up-regulation in highly adherent strains. As proof of principle and to investigate a possible role in pathogenesis for differentially expressed proteins, gain of function experiments were performed using two novel proteins that were among the most highly expressed surface proteins in adherent strains. Overexpression of either of these proteins, TVAG_244130 or TVAG_166850, in a relatively non-adherent strain increased attachment of transfected parasites to vaginal epithelial cells 2.2-fold. These data support a role in adhesion for these abundant surface proteins. Our analyses demonstrate that comprehensive profiling of the cell surface proteome of different parasite strains is an effective approach to identify potentialnew adhesion factors as well as other surface molecules that may participate in establishing and maintaining infection by this extracellular pathogen