Stach R W, Stach B M, Ennulat D J
Biochem Biophys Res Commun. 1986 Jan 29;134(2):1000-5. doi: 10.1016/s0006-291x(86)80520-4.
Dorsal root ganglia were extirpated from 9-day old embryonic chickens and solubilized in phosphate buffered saline containing 0.5% Noniodet P 40 detergent. When nerve growth factor binding studies are performed on these samples, the expected curvilinear Rosenthal (Scatchard) plot is obtained. However, when the solubilized cell sample is made 1-2 mM in phenylmethylsulfonyl fluoride and nerve growth factor binding is determined, a linear Rosenthal (Scatchard) plot is obtained. The equilibrium dissociation constant obtained from the slope of the line is 1.9 X 10(-9) M, identical to the equilibrium dissociation constant of the low affinity receptor. A similar phenomenon is observed when rat pheochromocytoma cells are solubilized in the non-ionic detergent and nerve growth factor binding is determined. No high affinity binding can be detected for either cell type when detergent solubilized cells are incubated with phenylmethylsulfonyl fluoride.
从9日龄胚胎鸡中取出背根神经节，将其溶解于含有0.5% Noniodet P 40去污剂的磷酸盐缓冲盐水中。当对这些样品进行神经生长因子结合研究时，可得到预期的曲线型罗森塔尔（斯卡查德）图。然而，当将溶解的细胞样品用苯甲基磺酰氟处理至1 - 2 mM并测定神经生长因子结合时，可得到线性的罗森塔尔（斯卡查德）图。从直线斜率获得的平衡解离常数为1.9×10⁻⁹ M，与低亲和力受体的平衡解离常数相同。当将大鼠嗜铬细胞瘤细胞溶解于非离子去污剂中并测定神经生长因子结合时，也观察到类似现象。当用苯甲基磺酰氟孵育经去污剂溶解的细胞时，两种细胞类型均检测不到高亲和力结合。
