Structural aspects and activation mechanism of human secretory group IIA phospholipase

Phospholipases are important probes for understanding structure–function relationships of membrane proteins. Many neurotoxins have phospholipase activity, and they have been recognized to be potential therapeutic agents for biological warfare. Understanding the modes of action of these enzymes is important for the development of effective therapeutic strategies. Human secretory phospholipases A2 (sPLA 2 ) interact with cellular membranes and catalyze the hydrolysis of phosphate ester bonds of phospholipids. The activity of these enzymes increases tremendously upon binding to a hydrophobic interface. Using molecular dynamics (MD) simulations in implicit solvent and membrane environments, we investigated alterations in structure and conformation of human sPLA 2 upon its interaction with a membrane that may be associated with the activation of the enzyme. In 50 ns MD simulations, starting from six different initial orientations of the protein relative to the membrane surface, the enzyme consistently adopted a membrane-bound configuration in close agreement with the known experimental data. The simulations also reproduced the experimentally determined distribution of hydrophobic and polar side chains on the interfacial binding surface. Differences in the dynamic behavior of the enzyme between the solvent and membrane-bound states were observed. In nonpolar media, the enzyme underwent major conformational rearrangements, which exposed the active site to the membrane. The increased mobility of the surface loop and the β-wing regions is required for the conformational change, which is essentially induced by the movement of N-terminal helix. Several active site residues underwent structural changes that reorganize the binding site for substrate catalysis. Overall, the results provided a valuable insight into the interfacial behavior of sPLA 2 enzyme and suggested that membrane binding is essential but insufficient for sPLA 2 activation.