The Amino Acid Sequence of Zinc-Carboxypeptidase from Streptomyces griseus

Abstract

The amino acid sequence of a zinc-carboxypeptidase from S. griseus (Cpase SG) was determined by automated Edman degradation and carboxypeptidase digestion of the S-carboxymethylated protein and by sequence analyses of peptides produced by cyanogen bromide cleavage and by lysyl endopeptidase digestion of the S-carboxymethylated protein. This enzyme is characterized by a uniquely broad substrate specificity which combines the specificities of mammalian Cpase A and Cpase B (J. Biochem. 86, 683-694, 1979). Cpase SG consists of 328 amino acid residues. The amino acid sequence of Cpase SG is partially similar to those of bovine Cpase A and Cpase B (sequence identity, 28-29%). In the sequence of Cpase SG, residues that are functionally important in mammalian Cpase A and Cpase B were all found at the corresponding positions. Residue 255 (according to the numbering system for bovine Cpase A), which, in the other Cpases, contributes to the difference in specificity between Cpase A (Ile-255) and Cpase B (Asp-255), was Asp. However, residue 254 was Ile, in contrast to Ser or Thr in all of the forms of Cpase A and Cpase B examined to date. The increase in hydrophobicity caused by the change at position 254 and the presence of negative charge at position 255 is probably one of the reasons for the broad substrate specificity of Cpase SG.