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J Biol Chem, Vol. 274, Issue 17, 11494-11504, April 23, 1999

Mechanisms by Which Elevated Intracellular Calcium Induces S49 Cell Membranes to Become Susceptible to the Action of Secretory Phospholipase A2

Heather A. WilsonDagger , Jacqueline B. WaldripDagger , Kelli H. NielsonDagger , Allan M. JuddDagger , Sang Kyou Han§, Wonhwa Cho§, Peter J. Sims, and John D. BellDagger

From the Dagger  Department of Zoology, Brigham Young University, Provo, Utah 84602, the § Department of Chemistry, the University of Illinois, Chicago, Illinois 60607-7061, and the  Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California 92037

Exposure of S49 lymphoma cells to exogenous group IIA or V secretory phospholipase A2 (sPLA2) caused an initial release of fatty acid followed by resistance to further hydrolysis by the enzyme. This refractoriness was overcome by exposing cells to palmitoyl lysolecithin. This effect was specific in terms of lysophospholipid structure. Induction of membrane susceptibility by lysolecithin involved an increase in cytosolic calcium and was duplicated by incubating the cells with calcium ionophores such as ionomycin. Lysolecithin also activated cytosolic phospholipase A2 (cPLA2). Inhibition of this enzyme attenuated the ability of lysolecithin (but not ionomycin) to induce susceptibility to sPLA2. Lysolecithin or ionomycin caused concurrent hydrolysis of both phosphatidylethanolamine and phosphatidylcholine implying that transbilayer movement of phosphatidylethanolamine occurred upon exposure to these agents but that susceptibility is not simply due to exposure of a preferred substrate (i.e. phosphatidylethanolamine) to the enzyme. Microvesicles were apparently released from the cells upon addition of lysolecithin or ionomycin. Both these vesicles and the remnant cell membranes were susceptible to sPLA2. Together these data suggest that lysolecithin induces susceptibility through both cPLA2-dependent and -independent pathways. Whereas elevated cytosolic calcium was required for both pathways, it was sufficient only for the cPLA2-independent pathway. This cPLA2-independent pathway involved changes in cell membrane structure associated with transbilayer phospholipid migration and microvesicle release.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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