IN SILICO STUDY OF ECHINOCOCCUS GRANULOSUS GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE ASSOCIATED WITH BISPHOSPHONATE

AGUERO F; ANDREA MAGLIOCO; PAULINO M; ALEJANDRA JUÁREZ VALDEZ; ROLDÁN, EMILIO AJ; FUCHS A

Congreso; Reunión Anual de Sociedades de Biociencias 2023; 2023

The Echinococcus granulosus (Eg) produces cystic echinococcosis, a zoonotic disease, in ungulate animals and in humans. Argentina reported 630 human cases in 2018-2019. In our laboratory we work using a cellular line from Eg G1 protoscolices, EGPE. The bisphosphonates (BF) on EGPE decreased cell growth and ATP, increased the total calcium but decreased the Ca2+. The ethidronate (EHDP) was effective and the alendronate (AL) had no effect. We identified the Eg?s glyceraldehyde-3-phosphate-dehydrogenase (GAPDH), a tetramer, UniProt W6UJ19 by MS/MS. We investigated whether GAPDH, could be druggable with EHDP and AL. Materials and Methods: A validated GAPDH model bound to NAD+, substrate, Pi and BF was subjected to molecular dynamics simulation (MD) for 100 ns in NAMD2. Structures were analyzed and visualized with software VMD 1.9.3 and MOE2022. Potential energy, RMSD and RMSF from the system with or without BF (control) were calculated. Binding free energy with ligands was calculated by MMPBSA in the last 100 frames of the MD. T-test was used to compare control and samples. Results: The EHDP was placed close to the Pi site and did not allow the Pi binding and the AL interacted with Thr154, Gln211, Thr 212, Gly213 and Arg235 impeding the substrate binding. Binding free energy for EHDP and GAPDH was favorable only in monomers A and B. The control showed favorable interaction with substrate only in monomer A (-10.7±5.1 Kcal/mol), while in presence of EHDP, the substrate had favorable interaction with 3 monomers (A, C, D). Significant differences were found between the binding free energy estimated for the substrate with each monomer with or without EHDP (p