Masuda Y, Ohtaki T, Kitada C, Tsuda M, Arimura A, Fujino M
Tsuruba Research Laboratories, Takeda Chemical Industries, Ltd., Ibaraki, Japan.
Biochem Biophys Res Commun. 1990 Oct 30;172(2):709-14. doi: 10.1016/0006-291x(90)90732-3.
A receptor for pituitary adenylate cyclase activating polypeptide (PACAP) was solubilized from bovine brain membranes with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate. Saturable binding experiments using [125I]PACAP27 revealed that the solubilized extract contained a single class of binding sites with a Kd of 200 pM and a Bmax of 0.9 pmol/mg protein. The Kd did not change significantly after the solubilization. Competitive binding experiments confirmed that the solubilized receptor retained a specificity for PACAP. Thus, the solubilization of the receptor in an active form was successfully achieved by the present procedure.
垂体腺苷酸环化酶激活多肽（PACAP）受体用3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸盐从牛脑膜中增溶出来。使用[125I]PACAP27进行的饱和结合实验表明，增溶提取物含有一类单一的结合位点，其解离常数（Kd）为200 pM，最大结合容量（Bmax）为0.9 pmol/mg蛋白质。增溶后Kd没有显著变化。竞争性结合实验证实，增溶的受体对PACAP保持特异性。因此，通过本方法成功实现了受体以活性形式的增溶。
