Primary structure of the Streptomyces R61 extracellular DD-peptidase. 2. Amino acid sequence data.
In order to confirm the Streptomyces codon usage, the Streptomyces R61 DD-peptidase was fragmented by cyanogen bromide cleavage of the carboxymethylated protein, trypsin digestion of the carboxymethylated protein and trypsin digestion of the protein treated with beta-iodopenicillinate and endoxo-delta 4-tetrahydrophthalic acid. The isolated peptides, which altogether represented more than 50% of the polypeptide chain, were sequenced. The data thus obtained were in excellent agreement with the primary structure of the protein as deduced from the nucleotide sequence of the cloned gene. Though a weak acylating agent, beta-iodopenicillanate reacted selectively with the active site of the DD-peptidase and formed an adduct which mas much more stable than that formed with benzylpenicillin, thus facilitating the isolation and characterization of the active-site peptide.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001665:Binding Sites; D003488:Cyanogen Bromide; D006868:Hydrolysis; D007700:Kinetics; D002267:Muramoylpentapeptide Carboxypeptidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D010397:Penicillanic Acid / Q000031:analogs & derivatives; D010446:Peptide Fragments / Q000302:isolation & purification; D010795:Phthalic Acids; D013302:Streptomyces / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000032:analysis; D014357:Trypsin
D000596:Amino Acids; D010446:Peptide Fragments; D010795:Phthalic Acids; D013438:Sulfhydryl Compounds; C051657:endoxo-delta(4)-tetrahydrophthalic acid; D010397:Penicillanic Acid; C031528:6-iodopenicillanic acid; D002267:Muramoylpentapeptide Carboxypeptidase; D014357:Trypsin; D003488:Cyanogen Bromide