Elucidating on the Quaternary Structure of Viper Venom Phospholipase A2 Enzymes in Aqueous Solution.

Abstract

This study focuses on the quaternary structure of the viper-secreted phospholipase A₂ (PLA₂), a central toxin in viper envenomation. PLA₂ enzymes catalyse the hydrolysis of the sn-2 ester bond of membrane phospholipids. Small-molecule inhibitors that act as snakebite antidotes, such as varespladib, are currently in clinical trials. These inhibitors likely bind to the enzyme in the aqueous cytosol prior to membrane-binding. Thus, understanding its controversial solution structure is key for drug design. Crystal structures of PLA₂ in the PDB show at least four different dimeric conformations, the most well-known being "extended" and "compact". This variability among enzymes with >50% sequence identity raises questions about their transferability to aqueous solution. Therefore, we performed extensive molecular dynamics (MD) simulations of several PLA₂ enzymes in water to determine their quaternary structure under physiological conditions. The MD simulations strongly indicate that PLA₂ enzymes adopt a "semi-compact" conformation in cytosol, a hybrid between extended and compact conformations. To our knowledge, this is the first study that determines the most favorable dimeric conformation of PLA₂ enzymes in solution, providing a basis for advancements in snakebite envenoming treatment. Recognizing snakebite envenoming as a neglected tropical disease has driven the search for efficient, affordable alternatives to the current antivenoms. Therefore, understanding the main drug targets within snake venom is crucial to this achievement.