Regulation of eDHFR-tagged proteins with trimethoprim PROTACs.
Regulation of eDHFR-tagged proteins with trimethoprim PROTACs.
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DOI：
10.1038/s41467-023-42820-3
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发表时间：
2023-11-03
影响因子：
16.6
通讯作者：
Sellmyer, Mark A.
中科院分区：
文献类型：
--
作者：
Etersque, Jean M.;Lee, Iris K.;Sharma, Nitika;Xu, Kexiang;Ruff, Andrew;Northrup, Justin D.;Sarkar, Swarbhanu;Nguyen, Tommy;Lauman, Richard;Burslem, George M.;Sellmyer, Mark A.
细胞和活体动物中蛋白质水平的时间控制可以用来提高我们对蛋白质功能的理解。此外，工程蛋白的控制可用于治疗应用。蛋白水解靶向嵌合体（PRoteolysis-TArgeting Chimeras, PROTACs）是一种小分子驱动的策略，通过招募E3连接酶到感兴趣的靶蛋白上，实现蛋白质丰度的快速、翻译后调节。在这里，我们通过将抗生素甲氧苄啶（TMP）与E3连接酶Cereblon的配体pomalidomide共价连接，开发了几种PROTAC分子。这些分子诱导目标蛋白（poi）的降解，这些蛋白在基因上融合到一个小的蛋白质结构域，即大肠杆菌二氢叶酸还原酶（eDHFR），这是TMP的分子靶点。我们发现各种edhfr标记的蛋白可以被PROTAC分子7c稳健地降解至95%的最大表达量。此外，通过蛋白质组学和生化分析，基于tmp的PROTACs对免疫调节亚胺药物（IMiD）敏感的新底物的表达影响最小。最后，我们展示了另一个已知的degron-PROTAC对的多重调控，以及在转移性癌症的啮齿动物模型中的可逆蛋白调控，证明了该系统的强大力量。总之，TMP PROTACs是体外和体内选择性和可逆降解edhfr标记蛋白的一种强大方法。控制细胞和动物中蛋白质的能力对实验研究很重要，并可能具有治疗应用。在这里，作者基于抗生素甲氧苄啶开发了一组新的异双功能小分子，可以降解带有大肠杆菌二氢叶酸还原酶（eDHFR）基因标记的蛋白质。
Temporal control of protein levels in cells and living animals can be used to improve our understanding of protein function. In addition, control of engineered proteins could be used in therapeutic applications. PRoteolysis-TArgeting Chimeras (PROTACs) have emerged as a small-molecule-driven strategy to achieve rapid, post-translational regulation of protein abundance via recruitment of an E3 ligase to the target protein of interest. Here, we develop several PROTAC molecules by covalently linking the antibiotic trimethoprim (TMP) to pomalidomide, a ligand for the E3 ligase, Cereblon. These molecules induce degradation of proteins of interest (POIs) genetically fused to a small protein domain, E. coli dihydrofolate reductase (eDHFR), the molecular target of TMP. We show that various eDHFR-tagged proteins can be robustly degraded to 95% of maximum expression with PROTAC molecule 7c. Moreover, TMP-based PROTACs minimally affect the expression of immunomodulatory imide drug (IMiD)-sensitive neosubstrates using proteomic and biochemical assays. Finally, we show multiplexed regulation with another known degron-PROTAC pair, as well as reversible protein regulation in a rodent model of metastatic cancer, demonstrating the formidable strength of this system. Altogether, TMP PROTACs are a robust approach for selective and reversible degradation of eDHFR-tagged proteins in vitro and in vivo. The ability to control proteins in cells and animals is important for experimental research and may have therapeutic applications. Here, the authors developed a new set of heterobifunctional small molecules based on the antibiotic trimethoprim that can degrade proteins that are genetically tagged with E. coli dihydrofolate reductase (eDHFR).
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影响因子：
3.5
作者：
BAKER, DJ;BEDDELL, CR;STAMMERS, DK
通讯作者：
STAMMERS, DK
影响因子：
16
作者：
Brownell, James E.;Sintchak, Michael D.;Dick, Lawrence R.
通讯作者：
Dick, Lawrence R.
影响因子：
4
作者：
Buckley DL;Raina K;Darricarrere N;Hines J;Gustafson JL;Smith IE;Miah AH;Harling JD;Crews CM
通讯作者：
Crews CM
影响因子：
48
作者：
Herm-Goetz, Angelika;Agop-Nersesian, Carolina;Meissner, Markus
通讯作者：
Meissner, Markus
影响因子：
3.7
作者：
Cho, Ukrae;Zimmerman, Stephanie M.;Wandless, Thomas J.
通讯作者：
Wandless, Thomas J.