Bacterial complementation as a means to test enzyme-ligand interactions.
Bacterial complementation as a means to test enzyme-ligand interactions.
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细菌互补作为测试酶-配体相互作用的一种手段。
DOI：
10.1007/s002530051274
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发表时间：
1998
影响因子：
5
通讯作者：
Eakin,AE
中科院分区：
文献类型：
--
作者：
Canyuk,B;Craig3rd,SP;Eakin,AE
建立了一种细菌互补试验，用于快速筛选大量化合物，以识别那些抑制酶靶标的化合物，用于基于结构的抑制剂设计。靶酶是次黄嘌呤磷酸核糖转移酶(HPRT)。这种酶已被提议作为抑制剂的潜在靶点，这些抑制剂可能被开发成治疗由几种寄生虫引起的疾病的药物。该筛选试验利用基因缺陷细菌，并辅之以在微量滴度平板中选择性培养的活性重组酶。通过比较在有和没有测试化合物的情况下生长的细菌的吸光度测量，可以快速地检测这些化合物对细菌生长的影响。抑制细菌生长的IC50值反映了化合物结合和/或抑制重组酶的能力。我们用来自恶性疟原虫和克氏锥虫的重组HPRT以及人类的酶对这种细菌互补筛选方法进行了测试。这些研究的结果表明，使用细菌补体选择的筛选试验可以用于鉴定以酶为靶标的化合物，并可以成为基于结构的药物设计工作的重要组成部分。
A bacterial complementation assay has been developed for the rapid screening of a large number of compounds to identify those that inhibit an enzyme target for structure-based inhibitor design. The target enzyme is the hypoxanthine phosphoribosyltransferase (HPRT). This enzyme has been proposed as a potential target for inhibitors that may be developed into drugs for the treatment of diseases caused by several parasites. The screening assay utilizes genetically deficient bacteria complemented by active, recombinant enzyme grown in selective medium in microtiter plates. By comparing absorbance measurements of bacteria grown in the presence and absence of test compounds, the effect of the compounds on bacterial growth can be rapidly assayed. IC50values for inhibition of bacterial growth are a reflection of the ability of the compounds to bind and/or inhibit the recombinant enzyme. We have tested this bacterial complementation screening assay using recombinant HPRT from the parasitesPlasmodium falciparumandTrypanosoma cruzi, as well as the human enzyme. The results of these studies demonstrate that a screening assay using bacterial complement selection can be used to identify compounds that target enzymes and can become an important part of structure-based drug design efforts.