Modulation of biosynthesis of phosphatidylcholine via CDP-choline in rat liver: influence of ethanol on the microsomal cholinephosphotransferase activity.
We have studied in vitro the effects of ethanol on the different enzymes involved in the biosynthesis of phosphatidylcholine (PC) via CDP-choline. Ethanol alters neither choline kinase (CK) nor CTP:phosphocholine cytidylyltransferase (CT) activities but, at levels higher than 50 mM, it does significantly inhibit microsomal cholinephosphotransferase (CPT) activity concomitantly with an increase in the ethanol concentration. A study of the kinetics of the reaction catalysed by CPT shows that ethanol decreases Vmax without altering Km, indicating a non-competitive inhibitory effect. An analysis of the thermodependence of CPT activity in the absence of ethanol reveals a break in the Arrhenius plot and thus a straight relationship between enzyme activity and the physico-chemical state of the microsomal membrane. Incubation of microsomes in the presence of ethanol increased the transition temperature from 25.8-28.2 degrees C. Microsomes were also incubated with n-alkanols with chain-lengths of fewer than five carbon atoms at concentrations which, according to their partition coefficients, produce equimolar levels in the membrane. Under these conditions all the alkanols caused the same inhibitory effect. All these results demonstrate that ethanol modulate the PC biosynthesis at the level of CPT activity and does not affect the CT enzyme. The inhibition found on CPT is clearly dependent on the alteration produced by ethanol on the hepatic microsomal membrane.
Department of Biochemistry and Molecular Biology, Faculty of Sciences, University of Granada, Spain.
M P
MP
Carrasco
J L
JL
Segovia
C
C
Marco
D000438:Alcohols / Q000494:pharmacology; D000818:Animals; D002797:Choline Kinase / Q000378:metabolism; D019916:Choline-Phosphate Cytidylyltransferase / Q000378:metabolism; D003566:Cytidine Diphosphate Choline / Q000378:metabolism*; D002798:Diacylglycerol Cholinephosphotransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000431:Ethanol / Q000494:pharmacology*; D007700:Kinetics; D008099:Liver / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013696:Temperature
D000438:Alcohols; D010713:Phosphatidylcholines; D000431:Ethanol; D003566:Cytidine Diphosphate Choline; D002797:Choline Kinase; D019916:Choline-Phosphate Cytidylyltransferase; D002798:Diacylglycerol Cholinephosphotransferase