Purification and characterization of PAMP‐12 (PAMP[9–20]) in porcine adrenal medulla as a major endogenous biologically active peptide

FEBS Letters

Kato, Johji¹  Ishiyama, Yuichiro¹  Eto, Tanenao¹  Kangawa, Kenji²  Washimine, Hisanori¹  Kitamura, Kazuo¹  Kuwasako, Kenji¹ 

[1] First Department of Internal Medicine, Miyazaki Medical College, Kihara, Kiyotake, Miyazaki 889-16, Japan;National Cardiovascular Center Research Institute, Fujishirodai, Suita, Osaka 565, Japan

关键词: Proadrenomedullin N-terminal 20 peptide;    Radioimmunoassay;    Distribution;    Characterization;    Peptide purification;    Hypotensive peptide;    PAMP;    proadrenomedullin N-terminal 20 peptide;    AM;    adrenomedullin;    ir-;    immunoreactive;    RIA;    radioimmunoassay;    RP-HPLC;    reversed-phase high performance liquid chromatography;    TFA;    trifluoroacetic acid;   

DOI  :  10.1016/S0014-5793(97)00971-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown