Photo-cross-linked small-molecule microarrays as chemical genomic tools for dissecting protein-ligand interactions
Photo-cross-linked small-molecule microarrays as chemical genomic tools for dissecting protein-ligand interactions
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DOI：
10.1002/asia.200600208
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发表时间：
2006-12-01
影响因子：
4.1
通讯作者：
Osada, Hiroyuki
中科院分区：
文献类型：
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作者：
Kanoh, Naoki;Asami, Aya;Osada, Hiroyuki
我们开发了一种独特的光交联方法，用于以不依赖于官能团的方式固定各种小分子。我们的方法取决于紫外线照射后从三氟甲基芳基二氮杂环丙烷产生的卡宾物种的反应性。在模型实验中证明，光生卡宾能够与每个测试的小分子反应，并且在大多数情况下它们产生多个缀合物。在阵列上固定化实验中还发现，各种小分子被固定化，并且固定化的小分子保留了它们与它们的结合蛋白相互作用的能力。利用这种方法，构建了约2000种天然产物和药物的光交联微阵列。发现这种光交联微阵列格式不仅可用于配体筛选，而且可用于研究结构-活性关系，即，通过利用光交联过程的非选择性性质，在小分子中发现的结构基序（或药效团）与其对蛋白质的结合亲和力之间的关系。
We have developed a unique photo-cross-linking approach for immobilizing a variety of small molecules in a functional-group-independent manner. Our approach depends on the reactivity of the carbene species generated from trifluoromethylaryldiazirine upon UV irradiation. It was demonstrated in model experiments that the photogenerated carbenes were able to react with every small molecule tested, and they produced multiple conjugates in most-cases. It was also found in on-array immobilization experiments that various small molecules were immobilized, and the immobilized small molecules retained their ability to interact with their binding proteins. With this approach, photo-cross-linked microarrays of about 2000 natural products and drugs were constructed. This photo-cross-linked microarray format was found to be useful not merely for ligand screening but also to study the structure-activity relationship, that is, the relationship between the structural motif (or pharmacophore) found in small molecules and its binding affinity toward a protein, by taking advantage of the nonselective nature of the photo-cross-linking process.