The amino acid sequences of carboxypeptidases I and II from Aspergillus niger and their stability in the presence of divalent cations.
The amino acid sequences of serine carboxypeptidase I (CPD-I) and II (CPD-II), respectively, from Aspergillus niger have been determined by conventional Edman degradation of the reduced and vinylpyridinated enzymes and peptides hereof generated by cleavage with cyanogen bromide, iodobenzoic acid, glutamic acid cleaving enzyme, AspN-endoproteinase and EndoLysC proteinase. CPD-I consists of a single peptide chain of 471 amino acid residues, three disulfide bridges and nine N-glycosylated asparaginyl residues, while CPD-II consists of a single peptide chain of 481 amino acid residues, has three disulfide bridges, one free cysteinyl residue and nine glycosylated asparaginyl residues. The enzymes are closely related to carboxypeptidase S3 from Penicillium janthinellum. Both Ca2+ and Mg2+ stabilize CPD-I as well as CPD-II, at basic pH values, Ca2+ being most effective, while the divalent ions have no effect on the activity of the two enzymes.
Svendsen
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I
Carlsberg Laboratory, Department of Chemistry, Gl. Carlsbergvej 10, DK-2500 Valby, Denmark. ibs@crc.dk
Dal Degan
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F
D000595:Amino Acid Sequence; D001234:Aspergillus niger / Q000201:enzymology*; D001426:Bacterial Proteins / Q000737:chemistry; D002118:Calcium / Q000494:pharmacology; D002268:Carboxypeptidases / Q000737:chemistry*; D002413:Cations, Divalent / Q000494:pharmacology; D004795:Enzyme Stability; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000737:chemistry; D008274:Magnesium / Q000494:pharmacology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D017433:Protein Structure, Secondary; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013696:Temperature
D001426:Bacterial Proteins; D002413:Cations, Divalent; D007527:Isoenzymes; D010446:Peptide Fragments; D002268:Carboxypeptidases; C022633:serine carboxypeptidase; D008274:Magnesium; D002118:Calcium