Role of Charge Properties of Bacterial Envelope in Bactericidal Action of Human Group IIA Phospholipase A 2 against Staphylococcus aureus *

Mammalian Group IIA phospholipases A 2 (PLA 2 ) potently kill Staphylococcus aureus. Highly cationic properties of these PLA 2 are important for Ca 2+ -independent binding and cell wall penetration, prerequisites for Ca 2+ -dependent degradation of membrane phospholipids and bacterial killing. To further delineate charge properties of the bacterial envelope important in Group IIA PLA 2 action against S. aureus, we examined the effects of mutations that prevent specific modifications of cell wall ( dltA ) and cell membrane ( mprF ) polyanions. In comparison to the parent strain, isogenic dltA − bacteria are ∼30–100× more sensitive to PLA 2, whereas mprF − bacteria are <3-fold more sensitive. Differences in PLA 2 sensitivity of intact bacteria reflect differences in cell wall, not cell membrane, properties since protoplasts from all three strains are equally sensitive to PLA 2. A diminished positive charge in PLA 2 reduces PLA 2 binding and antibacterial activity. In contrast, diminished cell wall negative charge by substitution of (lipo)teichoic acids with d -alanine reduces antibacterial activity of bound PLA 2, but not initial PLA 2 binding. Therefore, the potent antistaphylococcal activity of Group IIA PLA 2 depends on cationic properties of the enzyme that promote binding to the cell wall, and polyanionic properties of cell wall (lipo)teichoic acids that promote attack of membrane phospholipids by bound PLA 2.