Identification of a perchloric acid-soluble protein (PSP)-like ribonuclease from Trichomonas vaginalis
A perchloric acid-soluble protein (PSP), named here tv-psp1, was identified in Trichomonas vaginalis. It is expressed under normal culture conditions according to expressed sequence tag (EST) analysis. On the other hand, Tv-PSP1 protein was identified by mass spectrometry with a 40% of identity to human PSP (p14.1). Polyclonal antibodies against recombinant Tv-PSP1 (rTv-PSP1) recognized a single band at 13.5 kDa in total protein parasite extract by SDS-PAGE and a high molecular weight band analyzed by native PAGE. Structural analysis of Tv-PSP1, using dynamic light scattering, size exclusion chromatography, and circular dichroism spectroscopy, showed a trimeric structure stable at 7 M urea with 38% α-helix and 14% β-sheet in solution and a molecular weight of 40.5 kD. Tv-PSP1 models were used to perform dynamic simulations over 100 ns suggesting a stable homotrimeric structure. Tv-PSP1 was located in the nucleus, cytoplasm, and hydrogenosomes of T. vaginalis, and the in silico analysis by Search Tool for the Retrieval of Interacting Genes/Proteins (STRING) showed interactions with RNA binding proteins. The preliminary results of RNA degradation analysis with the recombinant Tv-PSP1 showed RNA partial deterioration suggesting a possible putative ribonuclease function.
KeywordsTrichomonas vaginalis Perchloric acid-soluble proteins Molecular dynamic Cellular localization Homotrimeric structure Size exclusion chromatographic
We acknowledge Biol. Anel Edith Lagunes Guillén, M.Sc. Laura Isabel Vazquez Carrillo, QFB Leticia Avila-González, and Brenda Herrera Villalobos for their technical assistance.
This work was supported by UACM and a grant from CONACyT (83808) Mexico (to M.E.A.S.). AVO was supported by a scholarship from ICyTDF (ICyTDF/SRI/70/2011).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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