Identification of a perchloric acid-soluble protein (PSP)-like ribonuclease from Trichomonas vaginalis.
Alma Villalobos-OsnayaGeorgina Garza-RamosIris N SerratosCésar Millán-PachecoArturo González-RoblesRossana ArroyoLaura Itzel Quintas-GranadosMaría Elizbeth Alvarez SánchezPublished in: Parasitology research (2018)
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.
Keyphrases
- mass spectrometry
- protein protein
- endothelial cells
- high resolution
- liquid chromatography
- binding protein
- gene expression
- small molecule
- molecular dynamics
- tandem mass spectrometry
- molecular docking
- high performance liquid chromatography
- molecular dynamics simulations
- bioinformatics analysis
- ms ms
- anti inflammatory
- solid state
- induced pluripotent stem cells
- plasmodium falciparum