Carboxylmethylation of insulin and glucagon in vitro

We studied the methyl acceptor capacity of insulin and glucagon in vitro. The levels of carboxylmethylation of pancreatic hormones (dpm x 10(3], when incubated with S-adenosyl-L-(3H-methyl)-methionine as methyl donor and purified protein carboxylmethylase, were: insulin (n = 6) 8.1 +/- 0.2 and 11.1 +/- 1.5 (mean +/- SEM) for 0.25 and 1.0 mg/ml, respectively; glucagon (n = 6) 17.0 +/- 3.2 and 40.2 +/- 2.5 (mean +/- SEM) for 0.5 and 1.0 mg/ml, respectively. On a molar basis, the methyl acceptor capacity was 1.0 dpm/pmol for insulin and 9.5 dpm/pmol for glucagon. Polyacrylamide gel electrophoresis of carboxylmethylated hormones showed a radioactivity (3H-methyl) peak that co-migrated with the corresponding 125I-hormone. Glucagon, but not insulin, seems to be a relatively good substrate for carboxylmethylation.