Induction of cytosolic Ca2+ elevation mediated by Mas-7 occurs through membrane pore formation

Abstract

Mas-7, a mastoparan derivative, induces elevation of intracellular free Ca2+ concentration ([Ca2+](i)) along two independent pathways. The minor contribution occurs via phospholipase C activation and is negatively regulated by treatment with phorbol 12-myristate 13-acetate, a protein kinase C activator. The major contribution involves plasma membrane pores allowing not only Ca2+, Mn2+, and Na+ to enter but also the uptake of ethidium bromide (314 Pa) and lucifer yellow (457 Da), but not fura-2 (831 Pa), Evans blue (961 Pa), and fluorescein-conjugate phalloidin (1,175 Da). Mas-7-induced current, as measured in planar lipid bilayers, reveals that Mas-7-induced pores have two slope conductances, 290 and 94 pS, and that the pores are nonselective for cations. The results also indicate that Mas-7 can produce pores by direct interaction with the plasma membrane without the involvement of membrane proteins and cytosolic factors. Besides in human neuroblastoma cells, similar Mas-7 effects were also observed in other cell lines such as HL-60, 1321N1 human astrocytoma, and bovine chromaffin cells. The data suggest that the Mas 7-induced [Ca2+](i) elevation is the combined result of Ca2+ release from stores via phosphoinositide turnover and prolonged Ca2+ influx through membrane pores.