Target Identification by Diazirine Photo-Cross-linking and Click Chemistry.
Target Identification by Diazirine Photo-Cross-linking and Click Chemistry.
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DOI：
10.1002/9780470559277.ch090167
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发表时间：
2009-12
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生物活性小分子的靶点鉴定通常是正向化学遗传学中的速率决定步骤。光亲和标记（PAL）代表了一个有用的生化策略，在复杂的蛋白质混合物中的目标识别。该单元描述了使用烷基二氮杂环丙烷为基础的光亲和探针和Cu（I）催化的点击化学共价标记和可视化的生物活性小分子的目标。还描述了用于鉴定蛋白质靶的探针修饰的蛋白质的亲和纯化的一般方法。
Target identification of biologically active small-molecules is often the rate-determining step in forward chemical genetics. Photo-affinity labeling (PAL) represents a useful biochemical strategy for target identification in complex protein mixtures. This unit describes the use of alkyl diazirine-based photo-affinity probes and Cu(I)-catalyzed click chemistry to covalently label and visualize the targets of biologically active small-molecules. A general method for affinity purification of probe-modified proteins, useful for identification of protein targets, is also described.