Tailoring protein nanomechanics with chemical reactivity.
Tailoring protein nanomechanics with chemical reactivity.
复制标题
DOI：
10.1038/ncomms15658
复制
发表时间：
2017-06-06
影响因子：
16.6
通讯作者：
Garcia-Manyes S
中科院分区：
文献类型：
--
作者：
Beedle AEM;Mora M;Lynham S;Stirnemann G;Garcia-Manyes S
弹性蛋白的纳米力学特性决定了各种组织的弹性。调节蛋白质延伸性的一种广泛的自然策略在于共价二硫键的存在，它显着增强蛋白质的硬度。形成二硫键的普遍体内策略需要专用酶的存在。在这里，我们提出了一种替代化学途径，通过基于天然存在的小分子的二硫键异构化来促进非酶氧化蛋白质折叠。使用单分子力钳光谱，在 DFT 计算和质谱测量的支持下，我们证明蛋白质半胱氨酸和攻击性低分子量硫醇之间的瞬时混合二硫键中间加合物的化学结构的细微变化对蛋白质的机械稳定性具有显着影响。这种方法提供了一个通用工具来合理化 S-硫醇化的动力学及其在调节蛋白质纳米力学中的作用，提供关于化学反应性如何调节蛋白质弹性的分子见解。翻译后修饰调节蛋白质的纳米力学。在这里，作者使用由密度泛函理论计算支持的单分子力钳光谱来展示反应性低分子量分子硫醇化合物如何直接影响机械蛋白质折叠。
The nanomechanical properties of elastomeric proteins determine the elasticity of a variety of tissues. A widespread natural tactic to regulate protein extensibility lies in the presence of covalent disulfide bonds, which significantly enhance protein stiffness. The prevalent in vivo strategy to form disulfide bonds requires the presence of dedicated enzymes. Here we propose an alternative chemical route to promote non-enzymatic oxidative protein folding via disulfide isomerization based on naturally occurring small molecules. Using single-molecule force-clamp spectroscopy, supported by DFT calculations and mass spectrometry measurements, we demonstrate that subtle changes in the chemical structure of a transient mixed-disulfide intermediate adduct between a protein cysteine and an attacking low molecular-weight thiol have a dramatic effect on the protein's mechanical stability. This approach provides a general tool to rationalize the dynamics of S-thiolation and its role in modulating protein nanomechanics, offering molecular insights on how chemical reactivity regulates protein elasticity. Post-translational modifications modulate nanomechanics of proteins. Here the authors use single-molecule force-clamp spectroscopy supported by density functional theory calculations to show how reactive low-weight molecular thiol compounds directly affect mechanical protein folding.