Purification of homologous protein carboxyl methyltransferase isozymes from human and bovine erythrocytes.
We have purified the two major isozymes of the L-isoaspartyl/D-aspartyl protein methyltransferase from both human and bovine erythrocytes. These four enzymes all have polypeptide molecular weights of approximately 26,500 and appear to be monomers in solution. Each of these enzymes cross-reacts with antibodies directed against protein carboxyl methyltransferase I from bovine brain. Their structures also appear to be similar when analyzed by dodecyl sulfate gel electrophoresis for the large fragments produced by digestion with Staphylococcus aureus protease V8 or when analyzed by high-performance liquid chromatography (HPLC) for tryptic peptides. The structural relatedness of these enzymes was confirmed by sequence analysis of a total of 433 residues in 32 tryptic fragments of the human erythrocyte isozymes I and II and of the bovine erythrocyte isozyme II. We found sequence identify or probable identity in 111 out of 112 residues when we compared the human isozymes I and II and identities in 127 out of 134 residues when the human and bovine isozymes II were compared. These results suggest that the erythrocyte isozymes from both organisms may have nearly identical structures and confirm the similarities in the function of these methyltransferases that have been previously demonstrated.
Gilbert
J M
JM
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
Fowler
A
A
Bleibaum
J
J
Clarke
S
S
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D007527:Isoenzymes / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011496:Protein Methyltransferases / Q000302:isolation & purification*; D011497:Protein O-Methyltransferase / Q000302:isolation & purification*; D012689:Sequence Homology, Nucleic Acid
D000596:Amino Acids; D007527:Isoenzymes; D011496:Protein Methyltransferases; D011497:Protein O-Methyltransferase