The presence of N-terminal pyroglutamyl residues in hog thyroglobulin.
A method was devised to isolate N-terminal peptide fragments from the polypeptide chains constituting thyroglobulin even in the case when the terminal amino groups are naturally blocked, for instance, acylated. Reduced and carboxymethylated hog thyroglobulin was first acetylated and digested with thermolysin. The blocked N-terminal peptide fragments were separated from the unblocked N-terminal fragments by column chromatography on Dowex 50, then on Dowex 1 after dinitrophenylation, and finally fractionated into ten fractions by paper chromatography after gel filtration on Sephadex G-10. Structural analyses by enzymic or partial acid hydrolysis of these peptide fractions failed to detect N-terminal acetyl amino acid. Instead, pyroglutamyl peptides including pyroglutamylleucine were found. By the same method, acetylated lysine and glycine were identified for chicken lysozyme and horse myoglobin, respectively. The use of thermolysin because of its unique specificity, and the possible relevance of the present result to the previous data on the N-terminal analysis of thyroglobulin are discussed.
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D002268:Carboxypeptidases; D004140:Dinitrophenols; D005971:Glutamates / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D011485:Protein Binding; D013552:Swine; D013820:Thermolysin; D013954:Thyroglobulin
D000596:Amino Acids; D004140:Dinitrophenols; D005971:Glutamates; D010446:Peptide Fragments; D013954:Thyroglobulin; D002268:Carboxypeptidases; D013820:Thermolysin