CAJ | 학술논문

帕金森病是一种常见的神经系统退行性疾病。多巴脱羧酶（ DDC）是帕金森病研究的靶点蛋白之一，但是目前没有高通量的测活模型。因此，需要构建一种高通量多巴脱羧酶抑制剂的筛选模型，用于发现新型抑制剂。采用克隆表达纯化得到多巴脱羧酶和用于酶偶联反应的磷酸烯醇式丙酮酸羧化酶（ PEPC）。基于一系列酶联反应将CO2固定，检测其含量，从而测定多巴脱羧酶的活性。结果得到人源多巴脱羧酶和磷酸烯醇式丙酮酸羧化酶的体外纯酶，建立了一种高通量筛选模型，并且从70个天然化合物中，筛选得到2个多巴脱羧酶的抑制剂。成功构建了一种基于体外纯酶高通量多巴脱羧酶抑制剂的筛选模型。
파금삼병시일충상견적신경계통퇴행성질병。다파탈최매（ DDC）시파금삼병연구적파점단백지일，단시목전몰유고통량적측활모형。인차，수요구건일충고통량다파탈최매억제제적사선모형，용우발현신형억제제。채용극륭표체순화득도다파탈최매화용우매우련반응적린산희순식병동산최화매（ PEPC）。기우일계렬매련반응장CO2고정，검측기함량，종이측정다파탈최매적활성。결과득도인원다파탈최매화린산희순식병동산최화매적체외순매，건립료일충고통량사선모형，병차종70개천연화합물중，사선득도2개다파탈최매적억제제。성공구건료일충기우체외순매고통량다파탈최매억제제적사선모형。
Parkinson′s disease is one of common nervous system degenerative diseases .Dopa decarboxylase ( DDC) has been thought as one of the drug target for Parkinson′s disease , but there is lacking of a high-throughput screening inhibitor model for DDC .To establish a high-throughput screening model for identification novel inhibitors of DDC , cloning, expression and purification of DDC and phos-phoenolpyruvate carboxylase ( PEPC) , monitoring the activity of DDC in the coupling reaction was studied .The CO2 decarboxylated by DDC was detected , based on a series of enzyme-linked reactions .Results showed that the pure enzymes of DDC and PEPC were ob-tained, and 2 inhibitors were found from 70 natural products using the screening model .