CONICET | Buscador de Institutos y Recursos Humanos

Aquaporins (AQP) constitute a well distributed and diversified protein family of channelswhose structure remain highly conserved throughout species. These channels transportmainly water but some of them also transport solutes such as glycerol, hydrogenperoxide, etc. Trypanosoma cruzi (Tc) would have different AQP variants in its genomebut there are only partial functional results for one of them. Thus, structuralcharacterization of TcAQP is key to evaluate the physiological relevance of thesechannels on the parasite and to get progress in the knowledge of structure-functionparticularities of this protein family. In this work, AQP compatible sequences weresearched for within Trytripdb database for all T cruzi strains. DNA was extracted fromtrypomastigotes (RA strain) cultured in Vero cell line. Each AQP sequence was elucidatedafter cloning in T7TS vector, bacteria transformation (E coli, DH5α), plasmid purificationand sequencing (Macrogen). Multiple sequence alignments of AQPs was performed usingall data base Tc strains and RA strain and a Phylogenetic analysis of all KinetoplastidAQPs was also performed. 4 TcAQP were found (α to δ) for 13 Tc strains. Protein sizes arearound 230 amino acids with the exception of β (450) and key amino acids and specifictransport related motifs were compatible with water specific transport (Frogger?spositions, Ar/R, NPA1 and NPA2). 3D models were built by ab initio methods (Rosetta,iTasser) and pores were characterized based on molecular dynamics results (AMBER14SBand LIPID14 force field; Hole). Conserved AQP like tetrameric structures were shown byall constructed 3D models. This work is the first step in the full characterization of allTcAQP, further functional analysis of the transport of each TcAQP in vitro is still needed.