Methylation of phospholipids in microsomes of the rat aorta.
The methylation of phospholipids by S-adenosyl-L-methionine was characterized in microsomes prepared from strips of rat aorta. In the presence of 0.5 microM S-adenosyl-L-methionine, endogenous phosphatidylethanolamine was methylated to form three products: phosphatidyl-N-monomethylethanolamine, phosphatidyl-N,N-dimethylethanolamine and phosphatidylcholine. In the presence of 150 microM S-adenosyl-L-methionine the methylation activity increased more than 50-fold and the principal radioactive product was phosphatidylcholine. Optimal activity was at pH 9 and no magnesium requirement was detected. Exogenous phosphatidylethanolamine, phosphatidyl-N-monomethylethanolamine and phosphatidyl-N,N-dimethylethanolamine served as substrates for the enzyme. The methylation of exogenous phosphatidyl-N,N-dimethylethanolamine proceeded at a slower rate. Incubation of trypsin with the aorta microsomes reduced the enzymatic activity and reduced the relative yield of phosphatidyl-N-monomethylethanolamine. Phospholipase C degraded the methylated phospholipids, but phosphatidyl-N,N-dimethylethanolamine appeared to be less accessible to the phospholipase. The phospholipid methylation activity was inhibited by the addition of S-adenosyl-L-homocysteine or by L-homocysteinethiolactone. When intact strips of rat aorta were incubated with L-[methyl-3H]methionine, [3H]methyl groups were incorporated into phospholipids. This incorporation was inhibited when L-homocysteinethiolactone was added to the incubation. Polarized fluorescence of diphenylhexatriene in aorta microsomes was measured to determine the apparent membrane fluidity. When intact strips of aorta were incubated with methionine or with L-homocysteinethiolactone, methionine enhanced and L-homocysteinethiolactone decreased apparent fluidity of the microsomal membranes. Phospholipid methylation activity was examined in aorta microsomes prepared from genetically spontaneous hypertensive SHR strain rats. Phospholipid methylation activity was substantially greater in the SHR aorta microsomes than in microsomes prepared from Wistar-Kyoto WKY control strain aorta. Membrane fluidity was greater in the SHR aorta microsomes than in the WKY aorta microsomes. The hypothesis that phospholipid methylation activity influences fluidity of membranes and the possible involvement of methylated phospholipids in aorta membrane functions are discussed.
D000818:Animals; D001013:Aorta, Thoracic / Q000378:metabolism*; D002855:Chromatography, Thin Layer; D005454:Fluorescence Polarization; D006710:Homocysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D008297:Male; D008745:Methylation; D008861:Microsomes / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012435:S-Adenosylhomocysteine / Q000494:pharmacology; D012436:S-Adenosylmethionine / Q000494:pharmacology; D014357:Trypsin / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism
D010743:Phospholipids; D006710:Homocysteine; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; C007957:homocysteine thiolactone; D010738:Type C Phospholipases; D014357:Trypsin