Biphasic changes in the sarcolemmal phosphatidylethanolamine N-methylation activity in catecholamine-induced cardiomyopathy

Phosphatidylethanolamine (PE) N-methylation activity was studied in rat heart sarcolemma at 1, 3, 9 and 24 h after an intraperitoneal injection of isoproterenol (40 mg/kg). Three reaction sites for PE N-methylation were examined by assaying the incorporation of radiolabeled methyl groups from S-adenosyl-L-methionine (AdoMet) into sarcolemmal PE molecules under optimal conditions. Total methylation activity at catalytic site I (studied by employing 0.055 microM AdoMet) was increased at 1 and 3 h after the isoproterenol injection and depressed at 24 h; 9 h samples showed no change. Similar biphasic alterations were seen for phosphatidyl-N-monomethylethanolamine, the major methylated product formed at site I. Alterations in the methylation activity at site I were associated with changes in Vmax values but the apparent affinity for AdoMet remained unaltered. No alterations were found in total methylation activities at sites II and III in isoproterenol treated preparations when studied by employing 10 and 150 microM AdoMet, respectively. An increase and a decrease in the PE N-methylation activity at site I were also observed in the sarcoplasmic reticular (microsomal) fraction from experimental hearts after 1 h and 24 h of the isoproterenol injection respectively, without changes at sites II and III. On the other hand, no changes were seen in the mitochondrial fraction. These results indicate biphasic alterations in the sarcolemmal and microsomal PE N-methylation activities during the development of catecholamine-induced cardiomyopathy.