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Volume 271, Number 51, Issue of December 20, 1996 pp. 32753-32759
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.

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Induction of Cytosolic Ca²⁺ Elevation Mediated by Mas-7 Occurs through Membrane Pore Formation

(Received for publication, April 26, 1996, and in revised form, September 18, 1996)

Byung-Chang Suh , Sook-Keun Song , Young-Kee Kim and Kyong-Tai Kim

From the Department of Life Science and Basic Science Research Institute, Pohang University of Science and Technology, Pohang, 790-784, Republic of Korea and the  Department of Agricultural Chemistry, Chungbuk National University, Cheongju, 361-763, Republic of Korea

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 Da) and lucifer yellow (457 Da), but not fura-2 (831 Da), Evans blue (961 Da), 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.

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