Browsing by Author "Silvennoinen, Meeri"

Malaria is a major cause of human mortality, morbidity, and economic loss. P. falciparum is one of six Plasmodium species that cause malaria and is widespread in sub-Saharan Africa. Many of the currently used drugs for malaria have become less effective, have adverse effects, and are highly expensive, so new ones are needed. mPPases are membrane integral pyrophosphatases that are found in the vacuolar membranes of protozoa but not in humans. These enzymes pump sodium ions and/or protons across the membrane and are crucial for parasite survival and proliferation. This makes them promising targets for new drug development. In this study we aimed to identify and characterize transient pockets in mPPases that could offer suitable ligand binding sites. P. falciparum was chosen because of its therapeutical interest, and T. maritima and V. radiata were chosen because they are test systems in compound discovery. The research was performed using molecular modelling techniques, mainly homology modelling, molecular dynamics, and docking. mPPases from three species were used to make five different systems: P. falciparum (apo closed conformation), T. maritima (apo open, open with ligand, and apo closed) and V. radiata (open with ligand). P. falciparum mPPase does not have a 3D structure available, so a homology model was built using the closest structure available from V. radiata mPPase as a template. Runs of 100 ns molecular dynamics simulations were conducted for these five systems: monomeric mPPase from P. falciparum and dimeric mPPases for the others. Two representative 3D structures for each of the five trajectories, the most dissimilar one to another, were selected for further analysis using clustering. The scrutinized 3D structures were first analyzed to identify possible binding pockets using two independent methods, SiteMap and blind docking (where no pre-determined cavity is set for docking). A second set of experiments using different scores (druggability, enclosure, exposure, …) and targeted docking were then run to characterize all the located pockets. As a result, only half of the catalytic pockets were identified. None of the transient pockets were identified in P. falciparum mPPase and all of them were located within the membrane. Docking was performed using compounds that have shown inhibiting behavior in previous studies but did not give good results in the tested structures. In the end none of the transient pockets were interesting for further study.