【摘要】 以³H-苄普地尔作为放射性配体,从牛脑中提取钙调素作为结合受体,采用平衡柱结合法.研究了³H-苄普地尔与牛脑钙调素的结合特性,建立了³H-苄普地尔放射受体结合分析法。³H-苄普地尔仅在Ca²⁺存在下才能与钙调素产生特异结合,结合具有饱和性和可逆性。Scatchard分析得K_D=2.52±0.44μmol/l,B_max=13.95±2.33nmol/mg蛋白,Hill分析得n_H=0.96±0.03。该方法的批内和批间变异系数均为10％,重现性较好。还介绍了使用本法于新设计合成的化合物的竞争性抑制试验初步结果。更多还原

【Abstract】 Using H-bepridil as a radioligand and calmodulin (CaM, an intracellular calcium receptor) as a receptor, the characteristics of 3H-bepridil binding to CaM from bovine brain were analyzed with equilibrium column binding technique. The 3H-bepridil radioreceptor binding assay is also established. Binding of 3H-bepridil to CaM is specific, saturable and reversible only in the presence of calcium. Scatchard analysis revealed that KD was 2.52±0.44 μmol/1 (n=5) and Bmax was 13.95±2.33 nmol/mg protein (n=5). Hill analysis demonstrated that nH was 0.96±0.03 (n=5). The intra-CV or inter-CV of this method was about .10% (n=22), indicating good stability and reproducibility. The results of competitive inhibition experiments of newly synthesized compounds indicates that the affinity of all bepridil analogs to CaM was similar, with that of bepridil.更多还原