Plasma membrane fractions from rat liver contain a phosphatidate phosphohydrolase distinct from that in the endoplasmic reticulum and cytosol.
Assays for two distinct phosphatidate phosphohydrolase activities were established based upon a differential inhibition by N-ethylmaleimide (NEM). The activity that is insensitive to this reagent in rat liver is predominantly in the plasma membrane fraction, whereas the NEM-sensitive activity is in the cytosolic and microsomal fractions. The NEM-insensitive activity is further distinguished from the NEM-sensitive phosphohydrolase by: (a) being relatively stable to heat; (b) not being inhibited by phenylglyoxal, butane-2,3-dione, cyclohexane-1,2-dione, 2,4-dinitrofluorobenzene, 7-chloro-4-nitrobenz-2-oxa-1,3-diazole, and diethyl pyrocarbonate; (c) being inhibited by NaF and phosphatidylcholine; and (d) not being stimulated by Mg2+. The NEM-insensitive activity was specific for phosphatidate. Both phosphohydrolase activities could be inhibited by chlorpromazine, propranolol, sphingosine, and spermine. The NEM-sensitive phosphatidate phosphohydrolase activity was increased by incubating hepatocytes for 12 h with glucagon and dexamethasone, and this effect was antagonized by insulin. The NEM-sensitive phosphohydrolase is concluded to be involved in glycerolipid synthesis. The activity of the NEM-insensitive phosphohydrolase was not altered by preincubation of rat hepatocytes in the short or long term with vasopressin, glucagon, insulin, triiodothyronine, or dexamethasone, but it might be modulated indirectly by sphingosine. The NEM-insensitive enzyme of the plasma membranes could be involved in signal transduction via the agonist-stimulated degradation of phosphatidylcholine through the phospholipase D pathway.
Jamal
Z
Z
Department of Biochemistry, Heritage Medical Research Centre, Edmonton, Alberta, Canada.
Martin
A
A
Gomez-Muñoz
A
A
Brindley
D N
DN
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured; D002746:Chlorpromazine / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000737:chemistry*; D005033:Ethylmaleimide / Q000494:pharmacology; D006358:Hot Temperature; D008099:Liver / Q000737:chemistry* / Q000166:cytology; D008274:Magnesium / Q000494:pharmacology; D008297:Male; D017830:Octoxynol; D010711:Phosphatidate Phosphatase / Q000037:antagonists & inhibitors / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000494:pharmacology; D011433:Propranolol / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D013096:Spermine / Q000494:pharmacology; D013110:Sphingosine / Q000494:pharmacology
D013096:Spermine; D011092:Polyethylene Glycols; D017830:Octoxynol; D011433:Propranolol; D010711:Phosphatidate Phosphatase; D008274:Magnesium; D013110:Sphingosine; D005033:Ethylmaleimide; D002118:Calcium; D002746:Chlorpromazine