title: Purification and characterization of PAMP-12 (PAMP[9-20]) in porcine adrenal medulla as a major endogenous biologically active peptide.
abstract: Proadrenomedullin N-terminal 20 peptide (PAMP-20) is a potent hypotensive peptide processed from the adrenomedullin (AM) precursor. We developed a specific radioimmunoassay which recognizes the C-terminal region of PAMP-20. Using this radioimmunoassay, the distribution of immunoreactive (ir-) PAMP was determined in porcine tissues. High concentrations of ir-PAMP were observed in the adrenal medulla and in the atrium, and these values were comparable to the corresponding concentrations of ir-AM. The concentration of ir-PAMP was almost the same as that of ir-AM in the kidney, while ir-PAMP was significantly lower than ir-AM in the ventricle, lung, and aorta. Reversed-phase high performance liquid chromatography in each porcine tissue sample revealed that two major peaks of ir-PAMP existed: one emerged at a position identical to that of authentic porcine PAMP-20; the other unknown peak was eluted earlier. The unknown peptide was purified to homogeneity from porcine adrenal medulla, and its complete amino acid sequence was determined. This peptide was found to be PAMP[9-20] with a C-terminal amide structure, and was named PAMP-12. Intravenous injections of PAMP-12 in anesthetized rats showed a significant hypotensive effect in a dose-dependent fashion, and the effect was comparable to that of PAMP-20. These data indicate that PAMP-12, a major component of ir-PAMP, is processed from the AM precursor, as is PAMP-20, and may participate in cardiovascular control.
authors.affiliation: First Department of Internal Medicine, Miyazaki Medical College, Kiyotake, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kuwasako
authors.forename: K
authors.initials: K
authors.lastname: Kitamura
authors.forename: Y
authors.initials: Y
authors.lastname: Ishiyama
authors.forename: H
authors.initials: H
authors.lastname: Washimine
authors.forename: J
authors.initials: J
authors.lastname: Kato
authors.forename: K
authors.initials: K
authors.lastname: Kangawa
authors.forename: T
authors.initials: T
authors.lastname: Eto
mesh_terms: D000313:Adrenal Medulla / Q000737:chemistry*; D053607:Adrenomedullin / Q000031:analogs & derivatives; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D007022:Hypotension / Q000139:chemically induced; D007668:Kidney / Q000737:chemistry; D008168:Lung / Q000737:chemistry; D009206:Myocardium / Q000737:chemistry; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D010455:Peptides / Q000032:analysis; D011506:Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D011863:Radioimmunoassay; D051381:Rats; D017421:Sequence Analysis; D013552:Swine; D014665:Vasodilator Agents / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology
chemical_list: D010446:Peptide Fragments; D010455:Peptides; D011506:Proteins; D014665:Vasodilator Agents; C108283:proadrenomedullin (9-20); D053607:Adrenomedullin