Investigating plasmepsin flexibility as a function of the flap region : a unique structural and dynamic feature of aspartic protease.

Abstract

Malaria is one of the most deadly infectious protozoan diseases known to man. It is spread by the Plasmodium parasite through the bite of the female Anopheles mosquito. Increasing resistance to currently available antimalarial drugs is a growing concern. Plasmepsins are malarial aspartic proteases, due to their characteristic mechanism of action, the fact that they are found in all Plasmodium species and are essential to parasitic survival they represent novel targets in the design of antimalarials. A unique structural feature of aspartic proteases and plasmepsins is the flap region lying perpendicular to the catalytic aspartic acid active, partially covering the active site. The flap region plays an important structural (and kinetic) role in regulating access to the active site, thereby regulating ligand binding. The present study focused on the flap dynamics of Plm I – V, proposing and validating parameters to accurately quantify the dynamic behaviour of the flap region. The catalytic aspartic acids is highly conserved in the plasmepsin family; sequence analysis revealed that although all plasmepsins are similar in structure, they differ greatly in the residues in the flap region. The heterogeneity in this region gives each plasmepsin unique substrate specificity and response to inhibitors. The parameters proposed in the present study gives a detailed account for the twisting of the flaps which move away from the active site in the absence of an inhibitor. Upon inhibitor binding, residues in the flap region form hydrogen bonds with the inhibitor pulling it inward towards the active site rendering the enzyme inactive. The parameters proposed in the present study will be of great value in the design of novel plasmepsin inhibitors, with increased efficacy and potency.