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引用次数: 0
摘要
BioPROTACs 是一种专为靶向降解蛋白质而设计的异功能蛋白质。虽然它们为调节疾病相关蛋白提供了潜在的治疗途径,但目前的策略都是静态的,缺乏动态调节蛋白降解的能力。在这里,我们介绍一种合成框架,利用蛋白酶控制开关对目标蛋白水平进行动态微调。我们的想法是利用蛋白酶作为外源输入和蛋白质降解之间的界面层,通过分离生物PROTACs上的两个结合域来调节目标蛋白质对E3连接酶的招募。通过将外部输入与主要蛋白酶层分离,只需对设计进行最小限度的修改,就能随时调整新的条件降解表型。我们利用两个高效的 "bioPROTAC "系统展示了这种方法的适应性:AdPROM和基于IpaH9.8的Ubiquibodies。利用 TEV 蛋白酶作为转换器,我们可以将小分子和光遗传输入连接起来,实现有条件的靶向蛋白质降解。我们的研究结果凸显了带有蛋白酶响应连接体的生物PROTACs作为条件性靶向蛋白质降解的多功能工具的潜力。
Protease-Responsive Toolkit for Conditional Targeted Protein Degradation
BioPROTACs are heterobifunctional proteins designed for targeted protein degradation. While they offer a potential therapeutic avenue for modulating disease-related proteins, the current strategies are static in nature and lack the ability to modulate protein degradation dynamically. Here, we introduce a synthetic framework for dynamic fine-tuning of target protein levels using protease control switches. The idea is to utilize proteases as an interfacing layer between exogenous inputs and protein degradation by modulating the recruitment of target proteins to E3 ligase by separating the two binding domains on bioPROTACs. By decoupling the external inputs from the primary protease layer, new conditional degradation phenotypes can be readily adapted with minimal modifications to the design. We demonstrate the adaptability of this approach using two highly efficient “bioPROTAC” systems: AdPROM and IpaH9.8-based Ubiquibodies. Using the TEV protease as the transducer, we can interface small-molecule and optogenetic inputs for conditional targeted protein degradation. Our findings highlight the potential of bioPROTACs with protease-responsive linkers as a versatile tool for conditional targeted protein degradation.
期刊介绍:
The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism.
Topics may include, but are not limited to:
Design and optimization of genetic systems
Genetic circuit design and their principles for their organization into programs
Computational methods to aid the design of genetic systems
Experimental methods to quantify genetic parts, circuits, and metabolic fluxes
Genetic parts libraries: their creation, analysis, and ontological representation
Protein engineering including computational design
Metabolic engineering and cellular manufacturing, including biomass conversion
Natural product access, engineering, and production
Creative and innovative applications of cellular programming
Medical applications, tissue engineering, and the programming of therapeutic cells
Minimal cell design and construction
Genomics and genome replacement strategies
Viral engineering
Automated and robotic assembly platforms for synthetic biology
DNA synthesis methodologies
Metagenomics and synthetic metagenomic analysis
Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction
Gene optimization
Methods for genome-scale measurements of transcription and metabolomics
Systems biology and methods to integrate multiple data sources
in vitro and cell-free synthetic biology and molecular programming
Nucleic acid engineering.