Sphingomyelin Inhibits Hydrolytic Activity of Heterodimeric PLA2 in Model Myelin Membranes: Pharmacological Relevance.

In this work, the heterodimeric phospholipase A₂, HDP-2, from viper venom was investigated for its hydrolytic activity in model myelin membranes as well as for its effects on intermembrane exchange of phospholipids (studied by phosphorescence quenching) and on phospholipid polymorphism (studied by ¹H-NMR spectroscopy) to understand the role of sphingomyelin (SM) in the demyelination of nerve fibers. By using well-validated in vitro approaches, we show that the presence of SM in model myelin membranes leads to a significant inhibition of the hydrolytic activity of HDP-2, decreased intermembrane phospholipid exchange, and reduced phospholipid polymorphism. Using AutoDock software, we show that the NH^δ+ group of the sphingosine backbone of SM binds to Tyr22(C=O_pb^δ–) of HDP-2 via a hydrogen bond which keeps only the polar head of SM inside the HDP-2's active center and positions the sn-2 acyl ester bond away from the active center, thus making it unlikely to hydrolyze the alkyl chains at the sn-2 position. This observation strongly suggests that SM inhibits the catalytic activity of HDP-2 by blocking access to other phospholipids to the active center of the enzyme. Should this observation be verified in further studies, it would offer a tantalizing opportunity for developing effective pharmaceuticals to stop the demyelination of nerve fibers by aberrant PLA₂s with overt activity – as observed in brain degenerative diseases – by inhibiting SM hydrolysis and/or facilitating SM synthesis in the myelin sheath membrane. Binding of sphingomyelin (SM) to catalytic subunits of HDP-2 predicted by Autodock. SM is given in sticks representation and HDP-2P is displayed in molecular surface (A) and line stick representation (B). For the stick diagrams, emerald represents carbon, blue–nitrogen, orange–phosphorus, red–oxygen, white–hydrogen. For molecular surface and lines, green represents carbon, blue–nitrogen, red–oxygen, white–hydrogen, yellow–sulfur. Yellow broken lines identify intermolecular bonds. Arrows point to sn-2 bonds. Hydrogen bond between NH^δ+ group and Tyr22(C=O_pb^δ–) and ionic bond between PO₄^– group and His48 drive the positioning of the sn-2 acyl ester bond away from the active center that suggests that SM binds non-productively in the active center of HDP-2 enzyme. [ABSTRACT FROM AUTHOR]