Bimodal distribution of phosphatidic acid phosphohydrolase in NG108-15 cells. Modulation by the amphiphilic lipids oleic acid and sphingosine.
The properties and bimodal distribution of phosphatidic acid phosphohydrolase (PAP) were investigated in neuroblastoma X glioma hybrid NG108-15 cells. Two PAP activities distinguished by their differential sensitivity to Mg2+ and Triton X-100 were identified in the cytosolic and microsomal fractions. A digitonin permeabilization method was employed to study the basal distribution of the cytosolic PAP and its redistribution upon cell exposure to amphiphilic lipids. Under conditions which release 100% of the cytosolic marker enzyme lactate dehydrogenase, only 60% of total cellular PAP activity was released into the medium through the digitonin-induced membrane pores, suggesting that about 40% of the total are membrane associated. Elevated plasma-membrane levels of phosphatidic acid, accomplished by incubating cells with Streptomyces chromofuscus phospholipase D, did not affect the distribution of cytosolic PAP. In contrast, oleic acid induced a marked concentration-dependent redistribution of the cytosolic enzyme to the particulate fraction. PAP redistribution was completely abolished in the presence of the sphingoid base sphingosine, previously shown to inhibit PAP activity in vitro (Lavie, Y., Piterman, O. & Liscovitch, M. (1990) FEBS Lett. 277, 7-10). Thus, the distribution of cytosolic PAP is reciprocally regulated by a long-chain (fatty) acid and a long-chain (sphingoid) base which are breakdown products of phospholipids and sphingolipids, respectively. These effects might influence PAP function in glycerolipid metabolism and signal transduction under physiological and pathophysiological conditions.
Aridor-Piterman
O
O
Department of Hormone Research, Weizmann Institute of Science, Rehovot, Israel.
Lavie
Y
Y
Liscovitch
M
M
D002413:Cations, Divalent; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003902:Detergents; D004072:Digitonin / Q000494:pharmacology; D008274:Magnesium / Q000378:metabolism; D017830:Octoxynol; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism*; D010711:Phosphatidate Phosphatase / Q000378:metabolism*; D010739:Phospholipase D / Q000378:metabolism; D011092:Polyethylene Glycols / Q000378:metabolism; D013110:Sphingosine / Q000378:metabolism*; D013302:Streptomyces / Q000201:enzymology; D014407:Tumor Cells, Cultured / Q000201:enzymology*
D002413:Cations, Divalent; D003902:Detergents; D009829:Oleic Acids; D019301:Oleic Acid; D011092:Polyethylene Glycols; D017830:Octoxynol; D010711:Phosphatidate Phosphatase; D010739:Phospholipase D; D008274:Magnesium; D004072:Digitonin; D013110:Sphingosine