Kanoh Naoki, Asami Aya, Kawatani Makoto, Honda Kaori, Kumashiro Saori, Takayama Hiroshi, Simizu Siro, Amemiya Tomoyuki, Kondoh Yasumitsu, Hatakeyama Satoru, Tsuganezawa Keiko, Utata Rei, Tanaka Akiko, Yokoyama Shigeyuki, Tashiro Hideo, Osada Hiroyuki
Antibiotics Laboratory, Discovery Research Institute, RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Chem Asian J. 2006 Dec 18;1(6):789-97. doi: 10.1002/asia.200600208.
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.
我们开发了一种独特的光交联方法，可独立于官能团固定各种小分子。我们的方法依赖于三氟甲基芳基二氮杂环丙烷在紫外线照射下产生的卡宾物种的反应性。模型实验表明，光生卡宾能够与测试的每个小分子反应，并且在大多数情况下会产生多种共轭物。在阵列固定实验中还发现，各种小分子被固定，并且固定的小分子保留了与它们的结合蛋白相互作用的能力。通过这种方法，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形式不仅可用于配体筛选，还可用于研究结构-活性关系，即通过利用光交联过程的非选择性，研究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。
