Department of Chemical Sciences, Tata Institute of Fundamental Research, Mumbai 400005, India.
Institute for Stem Cell Science and Regenerative Medicine, Bangalore 560065, India.
J Am Chem Soc. 2021 Nov 10;143(44):18766-18776. doi: 10.1021/jacs.1c09611. Epub 2021 Nov 1.
Protein-folding can go wrong and , with significant consequences for the living organism and the pharmaceutical industry, respectively. Here we propose a design principle for small-peptide-based protein-specific folding modifiers. The principle is based on constructing a "xenonucleus", which is a prefolded peptide that mimics the folding nucleus of a protein. Using stopped-flow kinetics, NMR spectroscopy, Förster resonance energy transfer, single-molecule force measurements, and molecular dynamics simulations, we demonstrate that a xenonucleus can make the refolding of ubiquitin faster by 33 ± 5%, while variants of the same peptide have little or no effect. Our approach provides a novel method for constructing specific, genetically encodable folding catalysts for suitable proteins that have a well-defined contiguous folding nucleus.
蛋白质折叠可能会出错，分别对生物体和制药行业产生重大影响。在这里，我们提出了一个基于小肽的蛋白质特异性折叠修饰物的设计原则。该原则基于构建“异核体”，即模拟蛋白质折叠核心的预折叠肽。我们使用停流动力学、NMR 光谱学、Förster 共振能量转移、单分子力测量和分子动力学模拟，证明异核体可以使泛素的重折叠速度提高 33±5%，而同一肽的变体几乎没有影响。我们的方法为具有明确连续折叠核心的合适蛋白质构建特定的、可遗传编码的折叠催化剂提供了一种新方法。
