Structural and phylogenetic analysis of Trypanosoma cruzi´s major intrinsic proteins (MIP)

TESAN FIORELA; LORENZO RAMIRO; GOREN, NORA BEATRIZ; ZERBETTO DE PALMA GERARDO; ZEIDA ARI; FOX ROMINA; ARMENTIA LUCIANO; PENAS FEDERICO; KARINA ALLEVA

Rockville

Congreso; Molecular Parasitology Meeting XXXI; 2020

Genetics Society of America

The rise in genome and transcriptome sequencing revealed a great diversity of aquaglyceroporins (Glp) and aquaporins (Aqp) (MIP, Major Intrinsic Protein superfamily) throughout the three domains of life: Archaea, Bacteria and Eukarya. Some of these channels, first identified as water transporters, also transport solutes such as glycerol, hydrogen peroxide, etc. Also, some of these channels are involved in the internalization of antiparasitic drugs in kinetoplastids: pentamidine for Trypanosoma brucei and trivalent antimony for Leishmania. Trypanosoma cruzi genome encode four Aqps (TcAqps) and only one of them was characterized (transport and localization). Thus, the aim of this work is to characterize the structure of TcAqps to gain knowledge on their structure-function particularities and to study their evolutionary context. Aqp compatible sequences were searched for within Trytripdb and NCBI databases (annotated as Aqp or resulted from tblastn search). Multiple sequence alignments (MAFFT, v7) were performed to build phylogenetic trees (Maximum Likelihood). Synteny studies were performed for those Trytripdb retrieved sequences. 3D models were built for the four TcAqps (Brener NEL stain) by ab initio methods (Rosetta, iTasser) and pores were characterized based on molecular dynamic analyses (AMBER14SB and LIPID14 force field; Hole). Conserved AQP like tetrameric structures were found in all constructed 3D models. MIP asymmetry was found to be the distinctive feature between american and african trypanosomas (not only T. brucei and T. cruzi) with the exception of T. grayi. For most of the other Trypanosomatida genera we found both Aqp and Glp MIPs. Aqps coding regions were lost in african trypanosomes within syntenic blocks. From the key MIP residues analysis, Trypanosomatida Aqpsshowed a classical Aqp profile but had a nonclassical selectivity filter. Trypanosomatida Glps showed a classical Glp and selectivity filter residue profiles. This work introduces a deeper understanding of the MIP superfamily and may help to elucidate its role in the Trypanosomatida order. Further functional analysis of the transport and localization of each TcAqp is still needed.