为高通量筛选和分子胶发现而优化的工程脑龙

IF 6.6 1区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Henry J. Bailey, Jonathan Eisert, Rubina Kazi, Jan Gerhartz, Dominika Ewa Pieńkowska, Ina Dressel, Joshua Vollrath, Ivan Kondratov, Tetiana Matviyuk, Nataliya Tolmachova, Varun Jayeshkumar Shah, Giulio Giuliani, Thorsten Mosler, Thomas M. Geiger, Ana M. Esteves, Sandra P. Santos, Raquel L. Sousa, Tiago M. Bandeiras, Eva-Maria Leibrock, Ulrike Bauer, Ivan Dikic
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引用次数: 0

摘要

大多数临床降解剂都使用一种基于免疫调节亚胺药物(IMiD)的衍生物,该衍生物可将其目标导向 E3 连接酶受体脑龙(CRBN);然而,IMiD 分子胶底物的鉴定工作仍未得到充分探索。为了解决这个问题,我们设计了人类 CRBN 构建物,它保留了三元复合物形成的所有特征,同时允许在大肠杆菌中进行同源和低成本表达。对该构建体的广泛分析表明,它在结合活性和生产简易性方面 "两全其美"。接下来,我们设计了 "以烯胺为重点的 IMiD 库",并证明了该构建物在高通量筛选中的适用性,确定了具有高效力、配体效率和特异性的结合体。最后,我们将我们的构建物用于原理验证胶水筛选方法,以确定 IMiD 细胞相互作用组。结合我们的 IMiD 结合图谱,这里描述的方法应成为协助发现下一代 CRBN 粘合剂的宝贵工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

An engineered cereblon optimized for high throughput screening and molecular glue discovery

An engineered cereblon optimized for high throughput screening and molecular glue discovery
The majority of clinical degraders utilize an immunomodulatory imide drug (IMiD)-based derivative that directs their target to the E3 ligase receptor cereblon (CRBN); however, identification of IMiD molecular glue substrates has remained underexplored. To tackle this, we design human CRBN constructs, which retain all features for ternary complex formation, while allowing generation of homogenous and cost-efficient expression in E. coli. Extensive profiling of the construct shows it to be the “best of both worlds” in terms of binding activity and ease of production. We next designed the “Enamine focused IMiD library” and demonstrated applicability of the construct to high-throughput screening, identifying binders with high potency, ligand efficiency, and specificity. Finally, we adapt our construct for proof of principle glue screening approaches enabling IMiD cellular interactome determination. Coupled with our IMiD binding landscape the methods described here should serve as valuable tools to assist discovery of next generation CRBN glues.
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来源期刊
Cell Chemical Biology
Cell Chemical Biology Biochemistry, Genetics and Molecular Biology-Molecular Medicine
CiteScore
14.70
自引率
2.30%
发文量
143
期刊介绍: Cell Chemical Biology, a Cell Press journal established in 1994 as Chemistry & Biology, focuses on publishing crucial advances in chemical biology research with broad appeal to our diverse community, spanning basic scientists to clinicians. Pioneering investigations at the chemistry-biology interface, the journal fosters collaboration between these disciplines. We encourage submissions providing significant conceptual advancements of broad interest across chemical, biological, clinical, and related fields. Particularly sought are articles utilizing chemical tools to perturb, visualize, and measure biological systems, offering unique insights into molecular mechanisms, disease biology, and therapeutics.
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