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用低浓度螯合剂乙二胺四乙酸(EDTA)去除肝细胞质膜的二价阳离子,继之以正丁醇抽提膜脂类,使胰岛素受体蛋白和其他一些膜蛋白进入水相,我们从肝脏得到不含任何去污剂的胰岛素受体蛋白.这样制备的胰岛素受体在Sephade xG-200凝胶层折中不滞留,经300,000×g超速离心70分钟不沉淀,且能通过0.22微米的微孔滤膜,从而证明它是完全水溶性的。胰岛素水溶性受体与胰岛素结合的解离常数Kd=3.6×10~(-9)M(24℃),该受体的等电点为pH 4.1,荧光疏水探针ANS不影响胰岛素与受体的结合,进一步证明在胰岛素与受体结合中,除疏水相互作用外,可能还存在其他的作用力。
With a low concentration of chelating agent ethylenediaminetetraacetic acid (EDTA) to remove the hepatic plasma membrane divalent cations, followed by n-butanol extracted membrane lipids, the insulin receptor protein and other membrane proteins into the water phase, we The liver was given an insulin receptor protein free of any detergent.The insulin receptor so prepared did not retain in the Sephadex xG-200 gel layer and was not precipitated by ultracentrifugation at 300,000 xg for 70 minutes and passed through a 0.22 micron Microporous membrane, which proves it is completely water-soluble. The dissociation constant of insulin-soluble insulin binding to insulin is 3.6 × 10 -9 M (24 ℃), the isoelectric point of this receptor is pH 4.1, and the fluorescent hydrophobic probe ANS does not affect the interaction between insulin and receptor , Further evidence of insulin binding to receptors, in addition to hydrophobic interactions, there may be other forces.