胆汁刀豆凝集素A结合蛋白在胆固醇结石早期形成阶段的作用

The role of biliary concanavalin A-binding proteins in the early stage of cholesterol gallstone formation: A comprehensive study of the nucleation activity

应用快速蛋白液相色谱分子筛与离子交换技术结合另一亲和层析系统——蜗牛凝集索(HP),对胆汁主要成核效应蛋白系统——刀豆凝集索A(ConA)结合蛋白的成核活性进行系统研究,得到一种抑成核的60KD蛋白与一种130KD的促成核蛋白。借助减法试验,对分离获得的这两种蛋白的成核重要性作了评估,为进一步寻找更重要的成核效应蛋白提供了又一途径。

As Con A-binding protein(CP) could reflect biliary nucleation activity, further isolation and purification of some proand anti-nucleating glycoproteins were condusted. CP was affinitively chromatographed by using Helix pomatia(HP)-Sepharose 6 MB, and then sieved through Superose 12 col umn connected to a fast protein liquid chromatography(FPLC) system. Based on nucleation activity study, i. e., the nucleation time (NT)and the final concentration of cholesterol monohydrate crystals, some effect-bearing molecular weight regions of CP were detected. We successfully isolated and purified a potent pro-nucleating glycoprotein with molecular weight of about 130 kD, and a potent anti-nucleating glycoprotein with molecular weight of 60 kD. By calculation based on the subtractive assay, we also found that the 130 kD glycoprotein accoundted for about 1/3 of total pro-nucleating activity of CP, and the 60 kD glycoprotein, 40% of HCP.