Effects of mastoparan B and its analogs on the phospholipase D activity in L1210 cells

Mastoparan B (MP-B), an amphiphilic alpha-helical peptide isolated from hornet venom, and its Ala-substituted analogs were examined for their effectiveness on phospholipase D (PLD) activity in L1210 cells. PLD activity was determined by measuring phosphatidylethanol produced from [3H]myristate-labelled cells in the presence of ethanol. PLD activity was stimulated by MP-B, 4MP-B (Lys4-->Ala), and 12MP-B (Lys12-->Ala), but not by 3MP-B (Leu3-->Ala) and 9MP-B (Trp9-->Ala). Other MPs including mastoparan 7 also stimulated the PLD activity, but inactive mastoparan 17 did not. The stimulatory effect of various MP analogs could be correlated with their alpha-helical contents. The PLD activity stimulated by MP-B was not affected by G-protein blocking chemicals. The extent of PLD stimulation by various MP-Bs, as well as by digitonin and beta-escin, correlated with the permeability of the membrane to ethidium bromide. These results suggest that the stimulation of PLD activity by MP-B in L1210 cells is probably coupled with membrane perturbation brought about by the peptide.