Forssmann K, Hock D, Herbst F, Schulz-Knappe P, Talartschik J, Scheler F, Forssmann W G
Klin Wochenschr. 1986 Dec 15;64(24):1276-80. 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.
一种新方法被应用于从人血液中分离一种多肽激素。来自1000升分子量低于20千道尔顿的血液滤过液中的多肽被吸附到2.5千克海藻酸上，然后洗脱、沉淀，并在G-25葡聚糖凝胶柱上脱盐，从而获得粗冻干血浆多肽提取物。这些多肽进一步进行离子交换色谱分析。此后，进行两步高效液相色谱法以纯化一种独特的多肽，在这种情况下它是心钠素（CDD）的循环形式。氨基酸分析、用羧肽酶A进行的C末端酶切以及序列分析表明，循环心钠素的唯一形式是含有28个氨基酸残基的分子，即从心钠素-126的C末端切割下来并与α-心钠肽（α心房钠尿肽）相同的心钠素-99-126。在血液滤过液中未检测到心钠素家族多肽激素的其他生物活性分子形式。分离过程之后是使用体外血管平滑肌舒张的生物测定法。
