Protein carboxy O-methylation is a kind of enzymatic reaction producing carboxy methylester catalyzed protein carboxyl O-methyltransferases at the carboxyl group of amino acid residues in polypeptide. Since the finding of carboxyl methylester, many st...
Protein carboxy O-methylation is a kind of enzymatic reaction producing carboxy methylester catalyzed protein carboxyl O-methyltransferases at the carboxyl group of amino acid residues in polypeptide. Since the finding of carboxyl methylester, many studies have been focused on the understanding of biological functions in eukaryotes but still not clear except for roles in Ras attachment to membrane and protein repair. In this study, we investigated the protein carboxyl methylation in porcine liver and testis in repect of identification and characterization of carboxyl methylesters and natural proteinous substrates using pH stability of the esters and electrophoresis under acidic and bacis conditions. We detected several kinds of methyl esters, 3 kimds each in cytosolic fractions from liver and testis. Under the treatment of strong acid and base, the ratio between base- stable substrates and unstable ones in liver (4:6) was different from the ratio obtainde in testis (6:4). The methyl accepting capacities were affected by enzymatic proteolysis between the range of 55 to 65% in liver and of 35 to 45% in testis. Separation of the methylated proteins by acidic elelctrophoresis in the presesnce of urea and SDS revealed distinctively natural substrates of 26, 33 and 80kD in the cytosol from liver, and of 14, 25, 32 and 86kD from testis. Most of the labelling, however, were lost following electrophoreses under moderare alkaline condition, except fot molecules of newly detected 7 and 17kD in liver, and 15, 29, 40 and 80kD in testis. From these results, it was proposed that protein carboxyl O-methylation in each organs may be catalyzed by different classes of protein carboxyl O- methyltransferases. In addition, it is suggested that the protein carboxyl methylation in liver and testis may have different patterns in respect of natural substrates.