Engineering FKBP-Based Destabilizing Domains to Build Sophisticated Protein Regulation Systems.
Engineering FKBP-Based Destabilizing Domains to Build Sophisticated Protein Regulation Systems.
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DOI：
10.1371/journal.pone.0145783
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Eickholt BJ
中科院分区：
文献类型：
--
作者：
An W;Jackson RE;Hunter P;Gögel S;van Diepen M;Liu K;Meyer MP;Eickholt BJ
用小分子靶向蛋白质稳定性已经成为以快速、可扩展和可逆的方式控制蛋白质丰度的有效工具。该技术涉及用由小分子特异性控制的去稳定结构域（DD）标记感兴趣的蛋白质（POI）。然而，这种融合蛋白的成功构建可能受到DD表位与蛋白质的完全生物学功能所需的静电相互作用的功能干扰的限制。这种方法的另一个缺点是剩余的内源性蛋白质。在这里，我们将Cre-LoxP系统与先进的DD相结合，并产生了一种蛋白质调控系统，其中内源性蛋白质的丢失，在我们的情况下，肿瘤抑制因子PTEN，可以直接与条件微调的DD-PTEN偶联。这个新系统将巩固和扩展DD技术的使用，以精确控制活细胞和动物模型中的蛋白质功能。
Targeting protein stability with small molecules has emerged as an effective tool to control protein abundance in a fast, scalable and reversible manner. The technique involves tagging a protein of interest (POI) with a destabilizing domain (DD) specifically controlled by a small molecule. The successful construction of such fusion proteins may, however, be limited by functional interference of the DD epitope with electrostatic interactions required for full biological function of proteins. Another drawback of this approach is the remaining endogenous protein. Here, we combined the Cre-LoxP system with an advanced DD and generated a protein regulation system in which the loss of an endogenous protein, in our case the tumor suppressor PTEN, can be coupled directly with a conditionally fine-tunable DD-PTEN. This new system will consolidate and extend the use of DD-technology to control protein function precisely in living cells and animal models.