Hiroshima O, Sano Y, Yuzuriha T, Yamato C, Saito A, Okamura N, Uchiyama Y, Kimura S, Goto K
Tsukuba Research Laboratories, Eisai Co., Ltd., Ibaraki, Japan.
J Neurochem. 1988 Feb;50(2):480-5. doi: 10.1111/j.1471-4159.1988.tb02936.x.
The binding site for calcitonin gene-related peptide (CGRP) was solubilized with 3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate (CHAPS) in an active form from porcine spinal cord. 125I-labeled human alpha-CGRP (125I-CGRP) binding to the solubilized protein was determined by filtration using a GF/B glass filter. The maximal binding activity (approximately 60% of the crude membrane fraction) was obtained with 5 mM CHAPS. 125I-CGRP binding to the solubilized protein was of high affinity, saturability, and high specificity, having KD and Bmax values of 3.69 pM and 338 fmol/mg of protein, respectively. The binding activity was eluted in a single peak with a molecular mass of 400,000 daltons by gel filtration on TSK gel G4000SW. These results suggest that the solubilized protein may be responsible for the specific binding site.
用3-[(3-胆酰胺丙基)二甲基铵]-1-丙烷磺酸盐（CHAPS）从猪脊髓中以活性形式溶解降钙素基因相关肽（CGRP）的结合位点。使用GF/B玻璃滤器通过过滤测定125I标记的人α-CGRP（125I-CGRP）与溶解蛋白的结合。用5 mM CHAPS可获得最大结合活性（约为粗膜部分的60%）。125I-CGRP与溶解蛋白的结合具有高亲和力、饱和性和高特异性，KD和Bmax值分别为3.69 pM和338 fmol/mg蛋白。通过在TSK凝胶G4000SW上进行凝胶过滤，结合活性以单一峰洗脱，分子量为400,000道尔顿。这些结果表明，溶解的蛋白可能是特异性结合位点的原因。
