Redirecting the specificity of tripartite motif containing-21 scaffolds using a novel discovery and design approach.
Redirecting the specificity of tripartite motif containing-21 scaffolds using a novel discovery and design approach.
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使用新颖的发现和设计方法重定向三方基序的特异性。
DOI：
10.1016/j.jbc.2023.105381
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发表时间：
2023-12
影响因子：
4.8
通讯作者：
Spangler, Jamie B.
中科院分区：
文献类型：
--
作者：
VanDyke, Derek;Xu, Linda;Sargunas, Paul R.;Gilbreth, Ryan N.;Baca, Manuel;Gao, Changshou;Hunt, James;Spangler, Jamie B.
劫持泛素蛋白酶体系统以诱导靶向蛋白降解(TPD)已成为在翻译后水平靶向和破坏细胞内蛋白的一种有前途的治疗策略。基于小分子的TPD方法，如蛋白水解靶向嵌合体(PROTAC)和分子胶，已经显示出潜力，目前有几种药物正在进行临床试验。生物PROTAC(BioPROTACs)是由靶结合结构域和E3泛素连接酶组成的工程融合蛋白，已成为治疗TPD的一种补充方法。在这里，我们描述了一种新的进化和设计生物PROTAC的方法。具体地说，基于人Tn3的第三个纤维连接蛋白III型结构域(Tn3)的工程结合支架被安装到包含-21的E3连接酶三段基序(TRIM21)中，以重定向其降解特异性。这是通过选择针对两种与B细胞淋巴瘤相关的不同致癌蛋白-粘膜相关淋巴组织淋巴瘤转位蛋白1(MALT1)和胚胎外胚层发育蛋白(EED)的天真酵母展示的Tn3文库，并用我们进化的Tn3结构域取代TRIM21的天然底物结合结构域来实现的。得到的TRIM21-Tn3融合蛋白既保留了Tn3的结合特性，又保留了TRIM21的E3连接酶活性。此外，我们在细胞模型中证明了TRIM21-Tn3融合蛋白通过泛素蛋白酶体系统有效地降解了各自的靶蛋白。我们探索了结合结构域亲和力和E3连接酶利用率的影响，以深入了解有效的BioPROTAC设计的要求。总体而言，这项研究提出了一种通用的工程方法，可用于针对治疗靶点设计和设计基于TRIM21的生物PROTAC。
Hijacking the ubiquitin proteasome system to elicit targeted protein degradation (TPD) has emerged as a promising therapeutic strategy to target and destroy intracellular proteins at the post-translational level. Small molecule–based TPD approaches, such as proteolysis-targeting chimeras (PROTACs) and molecular glues, have shown potential, with several agents currently in clinical trials. Biological PROTACs (bioPROTACs), which are engineered fusion proteins comprised of a target-binding domain and an E3 ubiquitin ligase, have emerged as a complementary approach for TPD. Here, we describe a new method for the evolution and design of bioPROTACs. Specifically, engineered binding scaffolds based on the third fibronectin type III domain of human tenascin-C (Tn3) were installed into the E3 ligase tripartite motif containing-21 (TRIM21) to redirect its degradation specificity. This was achieved via selection of naïve yeast-displayed Tn3 libraries against two different oncogenic proteins associated with B-cell lymphomas, mucosa-associated lymphoid tissue lymphoma translocation protein 1 (MALT1) and embryonic ectoderm development protein (EED), and replacing the native substrate-binding domain of TRIM21 with our evolved Tn3 domains. The resulting TRIM21–Tn3 fusion proteins retained the binding properties of the Tn3 as well as the E3 ligase activity of TRIM21. Moreover, we demonstrated that TRIM21–Tn3 fusion proteins efficiently degraded their respective target proteins through the ubiquitin proteasome system in cellular models. We explored the effects of binding domain avidity and E3 ligase utilization to gain insight into the requirements for effective bioPROTAC design. Overall, this study presents a versatile engineering approach that could be used to design and engineer TRIM21-based bioPROTACs against therapeutic targets.
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影响因子：
25
作者：
Fan, Xuelai;Jin, Wu Yang;Lu, Jie;Wang, Jin;Wang, Yu Tian
通讯作者：
Wang, Yu Tian
DOI：
10.1073/pnas.1507534112
发表时间：
2015-08-11
影响因子：
11.1
作者：
Fletcher, Adam J.;Mallery, Donna L.;James, Leo C.
通讯作者：
James, Leo C.
影响因子：
5.8
作者：
Fulcher LJ;Macartney T;Bozatzi P;Hornberger A;Rojas-Fernandez A;Sapkota GP
通讯作者：
Sapkota GP
影响因子：
64.8
作者：
Banik SM;Pedram K;Wisnovsky S;Ahn G;Riley NM;Bertozzi CR
通讯作者：
Bertozzi CR
影响因子：
7.3
作者：
Dong, Guoqiang;Ding, Yu;Sheng, Chunquan
通讯作者：
Sheng, Chunquan