Phosphatidylcholine synthesis-related enzyme activities of bovine brain microvessels exhibit susceptibility to peroxidation.
In microvessels isolated from bovine brain, microsomal enzyme activities involved in phosphatidylcholine biosynthesis and degradation were determined. The microvessels possessed acyl-CoA:1-acyl-sn-glycero-3-phosphocholine (AT) and glycerophosphocholine phosphodiesterase (GroPChoPDE) activity at a higher level compared with bovine and rat brain or rat liver microsomes whereas they expressed CTP:phosphocholine cytidylyltransferase (CT) and choline phosphotransferase (CPT) activity at a lower level. Each enzyme has been characterized in terms of response to inhibitors or activators revealing properties very similar to those in brain and liver microsomes. In the homogenate prepared from t-butylhydroperoxide-treated microvessels (10 min exposure to 10 microM up to 1 mM concentrations), AT and CPT activities exhibited a significant dose-dependent inhibition. In contrast, GroPChoPDE activity was unaffected. CT was inhibited only at 1 mM concentration. Short treatment of microvessels with Fe2+ (20 microM)-ascorbate (0.25 mM) or 100 microM linoleate hydroperoxide did not have any effect on the activity of the four enzymes. Strong inhibition of all enzymes was noted when the linoleate hydroperoxide system was fortified by Fe2+ ions (100 microM). AT inactivation was also found when oxidized low density lipoprotein was preincubated with microvessels. On the other hand, oxidized LDL left unchanged CPT and GroPChoPDE activities whereas it promoted a slight stimulation of cytidylyltransferase activity. Overall, the results suggest a link between oxygen radical generation and the perturbation of the microvessel membrane structure in which the four enzymes are incorporated, coupled to a direct sulfhydryl protein modification.
Sipione
S
S
Institute of Biochemistry, Faculty of Medicine, University of Catania, Italy.
Lupo
G
G
Anfuso
C D
CD
Albanese
V
V
Alberghina
M
M
D000818:Animals; D001921:Brain / Q000098:blood supply*; D002417:Cattle; D002798:Diacylglycerol Cholinephosphotransferase / Q000378:metabolism; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D015227:Lipid Peroxidation; D008315:Malondialdehyde / Q000378:metabolism; D008833:Microcirculation / Q000201:enzymology*; D008861:Microsomes / Q000201:enzymology*; D008862:Microsomes, Liver / Q000201:enzymology; D009713:Nucleotidyltransferases / Q000378:metabolism; D009928:Organ Specificity; D010545:Peroxides / Q000494:pharmacology; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species / Q000494:pharmacology; D013045:Species Specificity; D020122:tert-Butylhydroperoxide
D004791:Enzyme Inhibitors; D010545:Peroxides; D010713:Phosphatidylcholines; D017382:Reactive Oxygen Species; D008315:Malondialdehyde; D020122:tert-Butylhydroperoxide; D009713:Nucleotidyltransferases; C022348:phosphatidate cytidylyltransferase; D002798:Diacylglycerol Cholinephosphotransferase; D010727:Phosphoric Diester Hydrolases; C020677:glycerophosphocholine phosphodiesterase