Isolation and structural analysis of the circulating human cardiodilatin (alpha ANP)

• Published in: Klinische Wochenschrift Vol. 64; no. 24; p. 1276
• Main Authors: Forssmann, K, Hock, D, Herbst, F, Schulz-Knappe, P, Talartschik, J, Scheler, F, Forssmann, W G
• Format: Journal Article
• Language: English
• Published: Germany 15.12.1986
• Subjects: Amino Acid Sequence; Atrial Natriuretic Factor - isolation & purification; Chromatography, High Pressure Liquid; Humans; Kidney Failure, Chronic - blood; Molecular Weight; Muscle Proteins - isolation & purification; Peptide Termination Factors - isolation & purification
• ISSN: 0023-2173
• DOI: 10.1007/BF01785708

A new method was applied to isolate a polypeptide hormone from human blood. The polypeptides from 1,000 1 of hemofiltrate with a molecular weight lower than 20 kDaltons were adsorbed to 2.5 kg alginic acid, then eluted, precipitated, and desalted on a G-25 Sephadex column, thus obtaining a crude lyophilised plasma polypeptide extract. These polypeptides were further submitted to ion-exchange chromatography. Thereafter, two steps of HPLC were carried out to purify a distinct polypeptide which was the circulating form of cardiodilatin (CDD) in this case. The amino acid analysis, C-terminal enzymatic cleavage by carboxypeptidase A, and sequence analysis showed that the only form of circulating cardiodilatin is the 28 amino acid residue containing molecule, cardiodilatin-99-126 cleaved from the C-terminus of cardiodilatin-126 and identical with alpha-ANP (alpha atrial natriuretic polypeptide). Other bioactive molecular forms of the polypeptide hormones of the cardiodilatin family were not detected in the hemofiltrate. The isolation procedure was followed up by a bioassay using in vitro vascular smooth muscle relaxation.