The Application of the Chemical Array for Biological Study
The Application of the Chemical Array for Biological Study
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DOI：
10.1007/978-1-60761-845-4_8
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发表时间：
2010-01-01
期刊：
影响因子：
--
通讯作者：
Osada, Hiroyuki
中科院分区：
文献类型：
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作者：
Miyazaki, Isao;Simizu, Siro;Osada, Hiroyuki
鉴定小分子与人类感兴趣的蛋白质之间的特定相互作用是化学生物学和药物开发的基本步骤。与特定蛋白质结合的小分子可以作为研究细胞中蛋白质功能和生物过程的工具。我们已经开发了一种有效的方法来获得新的人类蛋白质结合配体的化学阵列方法。我们的方法包括使用表达感兴趣蛋白的细胞裂解物与红色荧光蛋白（RFP）融合，并通过合并显示分析进行高通量筛选，从而从阵列实验中去除use阳性信号。为了演示对各种感兴趣的人类蛋白质进行大规模配体筛选，建立了GLORIA基因文库（RIKEN Osada实验室用于化学阵列分析的基因文库）。利用该系统平台，我们检测到了碳酸酐酶II的新型抑制剂。我们还表明，这种筛选方法不仅对感兴趣的蛋白质的配体筛选有用，而且对深入了解结构亲和关系（SARs）和“基于片段的方法”的研究也有用。传统的基于片段的配体发现已经通过几种技术得到证实，如核磁共振波谱、x射线晶体学和质谱。我们目前的初步研究片段为基础的方法结合分析使用化学阵列格式。
The identification of specific interactions between small molecules and human proteins of interest is a fundamental step in chemical biology and drug development. The small molecules that bind to specific proteins can be used as tools to study the functions of proteins and biological processes in cells. We have developed an efficient method to obtain novel binding ligands of human proteins by a chemical array approach. Our method includes the use of cell lysates that express proteins of interest fused with red fluorescent protein (RFP) and high-throughput screening by merged display analysis, which removes Use positive signals from array experiments. To demonstrate large-scale ligand screening for various human proteins of interest, the gene library GLORIA (Gene Library of Osada Laboratory at RIKEN for chemical array analysis) has been established. Using the systematic platform, we detected novel inhibitors of carbonic anhydrase II. We also have shown that this screening method is useful not merely for ligand screening of proteins of interest, but also for gaining insight into structure affinity relationships (SARs) and for studies of "fragment-based approach."Traditional fragment-based ligand discoveries have been demonstrated by using several technologies, such as NMR spectroscopy and X-ray crystallography and mass spectrometry. We present initial studies of fragment-based approach to binding assay by using the chemical array format.