Isolation and amino acid sequence of COOH-terminal fragments from the beta subunit of human choriogonadotropin

• Published in: The Journal of biological chemistry Vol. 252; no. 15; pp. 5386 - 5392
• Main Authors: Birken, S, Canfield, R E
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 10.08.1977; American Society for Biochemistry and Molecular Biology
• Subjects: Amino Acid Sequence; Amino Acids - analysis; Chorionic Gonadotropin; Macromolecular Substances; Peptide Fragments - analysis; Thermolysin
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)63360-0

The amino acid sequence of the unique COOH-terminal region of the beta subunit of human choriogonadotropin has been reinvestigated. The desialylated subunit was digested with thermolysin and a 27-residue peptide from positions 115 through 141 isolated in a high yield. Quantitative Edman sequence degradation of this peptide, of another peptide produced by thermolysin digestion containing residues 142 to 145, and of two tryptic peptides (residues 123 to 145, 134 to 145) has established that the amino acid sequence of this region is: (formula: see text). In addition, the positions of attachment of the carbohydrate moieties to serine residues was established by a direct procedure using alkaline elimination and 35S-labeled sulfite addition, which yields [35S]-cysteic acid residues at the site of a substituted serine. Carbohydrate side chains in the COOH-terminal region have been shown to exist at residues 121, 127, 132, and 138. These studies have also resulted in the development of improved methods for the purification of COOH-terminal peptides of the human choriogonadotropin beta subunit.