CTP:phosphocholine cytidylyltransferase in intestinal mucosa.
CTP:phosphocholine cytidylyltransferase is thought to be a rate-limiting enzyme in phosphatidylcholine synthesis. This enzyme has not been well studied in intestine. We found that activity was greater in the non-lipid stimulated state (cytosolic form of the enzyme) than any previous tissue investigated (2.7 nM/min per mg protein). On addition of lysophosphatidylethanolamine, the enzyme only increased in activity 2.4-fold which is less than any previously reported tissue on lipid stimulation. As compared to liver, the enzyme was resistant to inhibition by chlorpromazine (gut, 100% activity remaining at 80 microM; 14% in liver). Tetracaine and propranolol were found to be impotent as inhibitors of the intestinal enzyme. Octanol-water partitioning showed that both chlorpromazine and tetracaine were hydrophobic, propranolol was not. pKa studies demonstrated that at the reaction pH, chlorpromazine would be uncharged. Physiologic experiments in which de novo phosphatidylcholine synthesis was either stimulated by bile duct fistulization and triacylglycerol infusion or suppressed by including phosphatidylcholine in a lipid infusion demonstrated that the enzyme (cytosolic enzyme) responded by decreasing Vmax but that the Km remained the same. In sum, these studies suggest that CTP:phosphocholine cytidylyltransferase in intestine is unique as compared to other tissues and that its response to a physiological stimulus is counter to that which would be adaptive.
D000818:Animals; D001646:Bile / Q000378:metabolism; D002746:Chlorpromazine / Q000494:pharmacology; D019916:Choline-Phosphate Cytidylyltransferase; D003600:Cytosol / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000201:enzymology*; D008297:Male; D008861:Microsomes / Q000201:enzymology; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010713:Phosphatidylcholines / Q000502:physiology; D010743:Phospholipids / Q000494:pharmacology; D011433:Propranolol / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility; D013048:Specimen Handling; D013748:Tetracaine / Q000494:pharmacology; D014280:Triglycerides / Q000378:metabolism
D010713:Phosphatidylcholines; D010743:Phospholipids; D014280:Triglycerides; D013748:Tetracaine; D011433:Propranolol; D009713:Nucleotidyltransferases; D019916:Choline-Phosphate Cytidylyltransferase; D002746:Chlorpromazine