Purification and characterization of a peptide C-terminal alpha-amidating enzyme from porcine atrium.
In many peptide hormones and neuropeptides, the carboxy-terminal alpha-amide structure is essential in eliciting biological activity. Here we report the purification and characterization of an alpha-amidating enzyme from porcine atrium, in which a high concentration of alpha-amidating activity was detected. The enzyme was purified to homogeneity from the membrane fraction of porcine atria by five steps of chromatography, including an affinity chromatography using a Sepharose column coupled with a substrate, Tyr-Phe-Gly. The purified enzyme was found to be composed of a single polypeptide chain with an apparent molecular weight of 92,000. This enzyme converted several synthetic peptides with C-terminal glycine to the corresponding des-glycine peptide alpha-amides.
Kojima
M
M
Department of Biochemistry, Miyazaki Medical College.
Mizuno
K
K
Kangawa
K
K
Matsuo
H
H
D000595:Amino Acid Sequence; D000818:Animals; D002848:Chromatography, DEAE-Cellulose; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006899:Mixed Function Oxygenases; D008970:Molecular Weight; D009097:Multienzyme Complexes; D009206:Myocardium / Q000201:enzymology*; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D013552:Swine
D009097:Multienzyme Complexes; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D000587:Oxidoreductases Acting on CH-NH Group Donors