Subject(s)
Anilides/pharmacology , Leukotriene B4/biosynthesis , Linoleic Acids/pharmacology , Neutrophils/metabolism , Arachidonate Lipoxygenases , Arachidonic Acid , Arachidonic Acids/biosynthesis , Chromatography, High Pressure Liquid , Humans , In Vitro Techniques , Leukotriene B4/blood , Lipoxygenase/blood , Neutrophils/drug effects , Phosphatidylcholines/blood , Phosphatidylinositols/bloodABSTRACT
An enzyme that catalyses the three-step methylation of phosphatidylethanolamine to phosphatidylcholine as well as the methylation of fatty acids and that uses S-adenosylmethionine as the methyl donor has been purified about 200-fold from rat liver. Irradiation of the purified enzyme with a short-wavelength u.v. light in the presence of [methyl-3H]8-azido-S-adenosylmethionine followed by electrophoresis results in the incorporation of radioactivity into a single protein band of about 25 kDa. It is concluded that a single catalytic subunit catalyses the conversion of phosphatidylethanolamine into phosphatidylcholine and fatty acid methylation.
Subject(s)
Liver/enzymology , Affinity Labels , Animals , Azides , Chromatography, Gel , Chromatography, High Pressure Liquid , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Hydrogen-Ion Concentration , Methylation , Rats , S-Adenosylmethionine/analogs & derivativesABSTRACT
Blood from humans and experimental animals was examined for the presence of platelet-activating factor. The procedure included a rapid extraction of the samples and a purification of the lipid content by thin layer chromatography. The chemical characterization was performed by phospholipases' treatment and high performance liquid chromatography. Rats contained the highest concentrations of platelet-activating factor and rabbits the lowest. Five anephric patients undergoing support hemodialysis had undetectable blood levels and the same finding was observed in six rats in which surgical nephrectomy was performed. Based on these data we suggest: 1) Low amounts of platelet-activating factor can be present in blood under normal conditions. 2) Kidney tissue seems to play a role in the formation of the platelet-activating factor that can be detected in blood under these conditions.
Subject(s)
Kidney Failure, Chronic/blood , Platelet Activating Factor/analysis , Animals , Blood Pressure , Chromatography, High Pressure Liquid , Humans , Male , Phospholipases/metabolism , Rabbits , Rats , Rats, Inbred StrainsABSTRACT
Changes in isoprenaline-sensitive phospholipid methyltransferase were studied in hepatocytes isolated from juvenile, mature and adrenalectomized rats. Isoprenaline produced greater stimulation of cyclic AMP accumulation in juvenile and mature adrenalectomized rats than in mature animals. Similarly, isoprenaline stimulated phospholipid methyltransferase in juvenile and mature adrenalectomized rats but had no effect in mature animals. Isoprenaline-mediated activation of phospholipid methyltransferase in adrenalectomized rats was time- and dose-dependent. In hepatocytes isolated from adrenalectomized rats incubated with [Me-3H]methionine or [3H]-ethanolamine the addition of isoprenaline increased the amount of radioactivity incorporated into phosphatidylcholine. The activation by isoprenaline of phospholipid methyltransferase was abolished by the beta-blocker propranolol and by insulin. These results indicate that rat liver the occupation of functional beta-receptors causes a stimulation of phospholipid methylation. It is suggested that, as reported previously, cyclic AMP activates phospholipid methyltransferase.
Subject(s)
Adrenalectomy , Isoproterenol/pharmacology , Liver/metabolism , Phosphatidylcholines/biosynthesis , Aging , Animals , Cyclic AMP/metabolism , Ethanolamines/metabolism , In Vitro Techniques , Insulin/pharmacology , Liver/cytology , Liver/drug effects , Male , Methionine/metabolism , Methyltransferases/metabolism , Phosphatidyl-N-Methylethanolamine N-Methyltransferase , Phosphatidylethanolamine N-Methyltransferase , Propranolol/pharmacology , Rats , Rats, Inbred StrainsABSTRACT
Suspensions of cyclic AMP sensitive cells of Dictyostelium discoideum responded to a cyclic AMP pulse with increased methylation of a protein of molecular weight about 120,000 and increased phospholipid demethylation. Protein methylation reached its peak 15-30 sec after cyclic AMP addition. Phospholipid demethylation reached its maximum within 2 min and basal levels were recovered in 3 min. S-Adenosyl-L-methionine is probably the methyl donor. In vitro addition of 0.25 mM and 25 microM S-adenosyl-L-methionine to sonicated D. discoideum cells inhibited ATP-dependent 45Ca2+ uptake by 70% and 25%, respectively. Based on these lines of evidence we propose that protein and phospholipid methylation are involved in D. discoideum chemotaxis probably by regulation of intracellular Ca2+ movements.
