A homogenous luminescence assay reveals novel inhibitors for giardia lamblia carbamate kinase.
A homogenous luminescence assay reveals novel inhibitors for giardia lamblia carbamate kinase.
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DOI：
10.2174/1875397301206010093
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Herzberg O
中科院分区：
文献类型：
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作者：
Chen CZ;Southall N;Galkin A;Lim K;Marugan JJ;Kulakova L;Shinn P;van Leer D;Zheng W;Herzberg O
人类病原体蓝氏贾第鞭毛虫是一种厌氧原生动物寄生虫，可导致贾第虫病，这是世界上最常见的腹泻疾病之一。虽然有几种药物可用于治疗贾第鞭毛虫病，但据报道对这些药物的耐药性可能会增加。贾第鞭毛虫氨基甲酸激酶(GlCK)在贾第鞭毛虫的新陈代谢中起着至关重要的作用，在人类中没有同源物，因此它是抗贾第鞭毛虫药物开发的一个有吸引力的候选者。我们建立了一种发光酶偶联测定法，通过定量该酶产生的三磷酸腺苷的量来测定glCK的活性。该分析是均相的，并已被小型化为1536孔板格式。用这种方法对4,096种已知化合物进行中试筛选，得到的信号与碱基比为11.5倍，Z‘因子为0.8，而glCK抑制剂的命中率为0.9%。因此，该方法可用于大量化合物的高通量筛选，用于药物开发中抑制物的鉴定。
The human pathogen Giardia lamblia is an anaerobic protozoan parasite that causes giardiasis, one of the most common diarrheal diseases worldwide. Although several drugs are available for the treatment of giardisis, resistance to these drugs has been reported and is likely to increase. The Giardia carbamate kinase (glCK) plays an essential role in Giardia metabolism and has no homologs in humans, making it an attractive candidate for anti-Giardia drug development. We have developed a luminescent enzyme coupled assay to measure the activity of glCK by quantitating the amount of ATP produced by the enzyme. This assay is homogeneous and has been miniaturized into a 1536-well plate format. A pilot screen against 4,096 known compounds using this assay yielded a signal-to-basal ratio of 11.5 fold and Z’ factor of 0.8 with a hit rate of 0.9 % of inhibitors of glCK. Therefore, this Giardia lamblia carbamate kinase assay is useful for high throughput screening of large compound collection for identification of the inhibitors for drug development.