Chemical genetic control of protein levels: Selective in vivo targeted degradation
Chemical genetic control of protein levels: Selective in vivo targeted degradation
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DOI：
10.1021/ja039025z
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发表时间：
2004-03-31
影响因子：
15
通讯作者：
Crews, CM
中科院分区：
文献类型：
--
作者：
Schneekloth, JS;Fonseca, FN;Crews, CM
基因功能缺失分析是研究蛋白质功能的有力手段。然而，由于缺乏合适的遗传模型系统，一些细胞生物学问题很难使用传统的遗传策略来解决。在这里，我们提出了一个通用的策略，设计和合成的分子能够诱导降解选定的蛋白质在体内通过泛素-蛋白酶体途径。蛋白质印迹和荧光分析表明，在分别用包含FKBP 12配体和双氢睾酮的PROteolysis Targeting嵌合分子（PROTACS）处理后，两种不同的靶点丢失：与FK 506结合蛋白（FKBP 12）融合的绿色荧光蛋白（GFP）和与雄激素受体（AR）融合的GFP。这些是第一个在体内的例子，直接小分子诱导的招聘目标蛋白质的蛋白酶体降解后，除了培养的细胞。此外，PROTAC介导的蛋白质降解提供了一个通用的策略，以创建“化学敲除”，从而打开新的可能性，控制蛋白质功能。
Genetic loss of function analysis is a powerful method for the study of protein function. However, some cell biological questions are difficult to address using traditional genetic strategies often due to the lack of appropriate genetic model systems. Here, we present a general strategy for the design and syntheses of molecules capable of inducing the degradation of selected proteins in vivo via the ubiquitin-proteasome pathway. Western blot and fluorometric analyses indicated the loss of two different targets: green fluorescent protein (GFP) fused with FK506 binding protein (FKBP12) and GFP fused with the androgen receptor (AR), after treatment with PROteolysis TArgeting Chimeric moleculeS (PROTACS) incorporating a FKBP12 ligand and dihydrotestosterone, respectively. These are the first in vivo examples of direct small molecule-induced recruitment of target proteins to the proteasome for degradation upon addition to cultured cells. Moreover, PROTAC-mediated protein degradation offers a general strategy to create "chemical knockouts," thus opening new possibilities for the control of protein function.