Mutt V
Fed Proc. 1979 Aug;38(9):2309-14.
Twenty-five years ago we described an extraction procedure for porcine secretin in which the intestinal tissue is briefly boiled in water and then extracted with dilute acid at low temperature. Boiling in water, which inactivates proteolytic enzymes, does not extract secretin, and extraction with acid in the cold will minimize cleavage of acid labile peptide structures. This extraction procedure has formed the basis for the isolation not only of secretin but also of cholecystokinin-pancreozymin (CCK) and, in collaboration with other laboratories, of the vasoactive intestinal peptide (VIP), the gastric inhibitory peptide (GIP), and motilin. Recently it has been used for the isolation of an N-terminally extended somatostatin from intestinal tissue, and of a peptide, from both nonantral gastric and intestinal tissues, with gastrin-releasing and probably cholecystokinin-releasing properties. A technique has been worked out permitting the chemical analysis, in certain cases, of polypeptide hormones in the presence of other polypeptides, the polypeptide mixture being exposed to fragmentation conditions known to result in characteristic hormone fragments, which are then extracted and quantitated. The technique can also be useful for the isolation of previously unknown peptides by identifying fragments of such and tracing them back to their peptides of origin.
25年前，我们描述了一种猪促胰液素的提取方法，即先将肠组织在水中短暂煮沸，然后在低温下用稀酸提取。在水中煮沸可使蛋白水解酶失活，但不会提取促胰液素，而在低温下用酸提取可使酸不稳定肽结构的裂解降至最低。这种提取方法不仅是促胰液素分离的基础，也是胆囊收缩素-促胰酶素（CCK）分离的基础，并且与其他实验室合作，还是血管活性肠肽（VIP）、胃抑制肽（GIP）和胃动素分离的基础。最近，它已被用于从肠组织中分离N端延长的生长抑素，以及从非胃窦部胃和肠组织中分离一种具有促胃泌素释放以及可能具有胆囊收缩素释放特性的肽。已经开发出一种技术，在某些情况下，可在存在其他多肽的情况下对多肽激素进行化学分析，即将多肽混合物暴露于已知会产生特征性激素片段的裂解条件下，然后对这些片段进行提取和定量。该技术还可用于通过鉴定此类片段并追溯其来源肽来分离先前未知的肽。
