Abnormal synthesis of N-methylated phospholipids during calcium paradox of the heart.

Phosphatidylethanolamine (PtdEtn) N-methyltransferase activity that synthesizes phosphatidylcholine (PtdCho) via formation of methylated intermediates (phosphatidyl-N-monomethylethanolamine, PtdEtnMe and phosphatidyl-N,N-dimethylethanolamine, PtdEtnMe2) was comparatively studied in rat heart sarcolemmal (SL), sarcoplasmic reticular (SR) and mitochondrial fractions during Ca2+ paradox. Perfusion (5 min) with Ca(2+)-free medium followed by reperfusion (5 min) with Ca(2+)-containing medium produced a marked rise in resting tension without any recovery of contractile force. Methyltransferase catalytic sites I, II and III which synthesize PtdEtnMe, PtdEtnMe2 and PtdCho, respectively, were assayed by measuring the [3H] methyl group incorporation from 0.055, 10 and 150 microM S-adenosyl-L-[3H-methyl] methionine into membrane PtdEtn molecules. Five minutes of perfusion with Ca(2+)-free medium did not affect either SL or SR N-methyltransferase systems. Ca(2+)-readmission for 1 to 5 min induced a selective, time-dependent depression of SL site II and SR site I methyltransferase activities. Individual N-methylated phospholipids specifically formed at the two sites reflected these changes. The above abnormalities were differently influenced by the duration (1-5 min) of Ca(2+)-free perfusion and were characterized by different kinetic alterations. The mitochondrial methylation system was not affected under Ca2+ paradox. The results suggest that reduced synthesis of SL N-methylated phospholipids may contribute to the contractile dysfunction observed in Ca2+ paradox.