Méster J, Rbertson D M, Feherty P, Kellie A E
Biochem J. 1970 Dec;120(4):831-6. doi: 10.1042/bj1200831.
An assay method was developed for the determination of high-affinity oestradiol receptors in uterine supernatant preparations. When only high-affinity sites are present in such preparations, or when they predominate, the analysis of the equilibrium between oestradiol and receptor sites based on the Scatchard (1949) plot may be used to determine the dissociation constant of the equilibrium and the molar concentration of the high-affinity sites. When both high-affinity and low-affinity sites are present the Scatchard plot is no longer linear and cannot be used directly to determine high-affinity sites. Determination of the reverse velocity constants of the reaction between high-affinity (k(-1)) and low-affinity (k(-2)) receptor sites and [(3)H]oestradiol has shown that these constants differ by at least one order of magnitude. Advantage has been taken of this difference to introduce an additional step into the assay procedure that eliminates oestradiol bound to low-affinity sites and permits the determination of high-affinity sites in different species and under a variety of physiological conditions.
开发了一种用于测定子宫上清液制剂中高亲和力雌二醇受体的分析方法。当此类制剂中仅存在高亲和力位点或高亲和力位点占主导时，基于Scatchard（1949年）图对雌二醇与受体位点之间的平衡进行分析，可用于确定平衡的解离常数和高亲和力位点的摩尔浓度。当同时存在高亲和力和低亲和力位点时，Scatchard图不再呈线性，不能直接用于确定高亲和力位点。对高亲和力（k(-1)）和低亲和力（k(-2)）受体位点与[³H]雌二醇之间反应的逆向速度常数的测定表明，这些常数至少相差一个数量级。利用这一差异在分析过程中引入了一个额外步骤，该步骤可消除与低亲和力位点结合的雌二醇，并允许在不同物种和各种生理条件下测定高亲和力位点。
