Synthesis of phosphatidylcholine by two distinct methyltransferases in rat colonic brush-border membranes: evidence for extrinsic and intrinsic membrane activities.
The enzymatic synthesis of phosphatidylcholine from phosphatidylethanolamine via a transmethylation pathway has not been shown to occur in the small intestine and has been assumed to be absent from the entire gut. The existence of this pathway, however, has not been investigated in the large intestine. Utilizing a recently developed method for the isolation of brush-border membranes from rat colonocytes, the present studies were designed to determine whether phospholipid methylation activity was present in the large intestine. The results demonstrate that this pathway for synthesis of phosphatidylcholine exists in rat colonic plasma membranes and involves at least two distinct methyltransferases. The predominant product of the first enzyme (methyltransferase I) is phosphatidyl-N-monomethylethanolamine; phosphatidylcholine and phosphatidyl-N-monomethylethanolamine are the principal products of the second enzyme (methyltransferase II). Methyltransferase I has an apparent Km for S-adenosyl-L-methionine of 100.0 microM and a pH optimum of 8.0, while methyltransferase II has an apparent Km of 0.3 microM and a pH optimum of 6.0. Additional evidence to support the presence of two distinct enzymes includes the differential effects of ATP, Triton X-100, trypsin treatment, and temperature on their activities.
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D003106:Colon / Q000201:enzymology; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000201:enzymology*; D008297:Male; D008745:Methylation; D008780:Methyltransferases / Q000037:antagonists & inhibitors / Q000145:classification / Q000378:metabolism*; D008871:Microvilli / Q000201:enzymology; D017830:Octoxynol; D010713:Phosphatidylcholines / Q000096:biosynthesis*; D010743:Phospholipids / Q000378:metabolism; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats; D012435:S-Adenosylhomocysteine / Q000494:pharmacology; D012436:S-Adenosylmethionine / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology; D013696:Temperature; D013997:Time Factors
D002413:Cations, Divalent; D010713:Phosphatidylcholines; D010743:Phospholipids; D011092:Polyethylene Glycols; D012436:S-Adenosylmethionine; D000255:Adenosine Triphosphate; D017830:Octoxynol; D012435:S-Adenosylhomocysteine; D008780:Methyltransferases