亲和毛细管电泳法对单胺氧化酶与犬尿胺特异性相互作用的研究

摘要

采用亲和毛细管电泳技术对粗提单胺氧化酶(MAO)与其专一性底物犬尿胺(Kyn)的相互作用进行研究,并引入MAO抑制剂氯吉兰对该特异性相互作用进行验证.结果表明,随着缓冲液中MAO浓度的增大,Kyn相对两个内标的迁移时间比率(RMTR)逐渐减小.而加入氯吉兰后,Kyn的RMTR值比未加入抑制剂时有所增大.说明MAO活性位点被氯吉兰结合,导致MAO与Kyn间的相互作用被抑制.该方法在表征MAO与Kyn相互作用的同时,还可为MAO抑制剂的筛选提供重要依据.与传统的MAO抑制剂筛选方法分光光度法和化学放射法相比,该方法具有简便、快速、样品用量少的优点.%The interaction between monoamine oxidase (MAO) and substrate Kyn was characterized by affinity capillary electrophoresis. The specific interaction between them was further demonstrated by adding an inhibitor clorgiline to MAO. The results demonstrated that the relative migration time ratio (RMTR) of Kyn decreased with the increase of MAO concentration in running buffer and the chromatographic peak of Kyn migrated far from that of triethyl benzyl ammonium chloride (TEBA) gradually.However, after adding the MAO inhibitor clorgiline, it was found that the values of RMTR of Kyn increased. This showed that the active site of MAO was occupied by clorgiline and the interaction between Kyn and MAO was inhibited. The developed method can be used to characterize the specific interaction between MAO and Kyn. Compared with the traditional methods of MAO inhibitors screening, e.g. spectrophotometric assay and radiochemical analysis, the presented method could provide a potential analytical tool for the screening of MAO inhibitor with the advantages of rapidness, convenience and less sample consumption.