蛋白-脂质体复合物毛细管电泳筛选单胺氧化酶抑制剂

基于酶与底物间的相互作用,建立了蛋白-脂质体复合物毛细管电泳筛选单胺氧化酶(MAO)抑制剂的新方法.分别将不同浓度的4种N-炔丙基胺类化合物添加至含有蛋白-脂质体复合物的毛细管电泳缓冲液中,抑制MAO活性.考察MAO底物犬尿胺(Kyn)在含有不同浓度N-炔丙基胺类化合物缓冲液中的迁移时间比率(RMTR).结果表明,化合物N-炔丙基-N-甲基-R-2-庚胺(R-2-HMP)和N-炔丙基-R-2-庚胺(R-2-HPA)能够明显抑制MAO活性,导致MAO与Kyn的相互作用减弱,Kyn的RMTR值随着R-2-HMP和R-2-HPA的增加呈现出明显增加的趋势.化合物N,N-二炔丙基-R-2-己胺和N,N-二炔丙基-R-2-辛胺对MAO活性抑制不明显,Kyn的RMTR随这两种浓度增加变化不大.此结果与柱外孵育测定化合物活性结果一致.与传统MAO筛选剂筛选方法相比,本方法快速,成本低,酶消耗量少且不受分离电压等于扰因素的影响.

Screen of Monoamine Oxidase Inhibitors by Protein-Liposome Conjugate Capillary Electrophoresis

Based on the interaction between enzyme and substrate,a simple and elegant method was developed to screen the inhibitors of monoamine oxidase(MAO).Four kinds of N-propargyl amine compounds with gradient concentrations were added in the CE running buffer containing protein-liposome conjugate to inhibit the activity of MAO to some extent.The relative migration time ratio(RMTR) of kynuramine(Kyn) was investigated in running buffer containing different compounds in gradient concentrations.The results showed that compound 1 and 2 could inhibit the activity of MAO obviously,such that the interaction between MAO and Kyn was weakened,RMTR of Kyn increased with the increase of compound 1 and 2.The inhibition activities of compound 3 and 4 were not strong,and RMTR of Kyn changed little with the increase of compound 3 and 4.This result was consistent with that of activity assay of compounds incubation offline.In comparison with the traditional screening method for MAO inhibitors,this method is simple,rapid,less consumption of enzyme and did not suffer from the interference caused by non-target molecules.