一种测定功能蛋白与其配体结合参数的简单方法

A simple method to determine association parameters between functional proteins and drugs

以羰基二咪唑法分别将牛血清白蛋白(BSA)和β2-肾上腺素受体(β2-AR)键合在大孔硅胶上,合成两种色谱固定相。采用直接进样法测定BSA与奥美拉唑、普萘洛尔和盐酸异丙嗪的结合常数及结合位点数,β2-AR与盐酸异丙肾上腺素、沙丁胺醇和去甲肾上腺素的结合常数及结合位点数。方法可用于功能蛋白与其配体分子相互作用的研究。

The bovine serum albumin and β2 adrenoceptor ligands were immob-ilized on the surface of macroporous silica gel through 1,1′-carbonyldiimidazole activation method to prepare the stationary phase.The association constants and binding sites of three kinds of drugs,omeprazole,propranolol and promethazine on BSA-bond medium,and salbutamol,isoprenaline and noradrenaline on β2-AR-bond medium were determined by altering their injected amount,respectively.The values of K and na were confirmed to be consistent with the results by spectrofluorometric method and reported results in the literatures,indicating that the proposed method can be used to investigate the interaction between ligands and functional proteins.