University of Victoria -Genome British Columbia Proteomics Centre, #3101-4464 Markham Street, Vancouver Island Technology Park, Victoria, BC V8Z7X8, Canada.
Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, NC 27599, USA.
J Proteomics. 2020 Jan 16;211:103544. doi: 10.1016/j.jprot.2019.103544. Epub 2019 Nov 1.
For disordered proteins, ligand binding can be a critical event that changes their structural dynamics. The ability to characterize such changes would facilitate the development of drugs designed to stabilize disordered proteins, whose mis-folding is important for a number of pathologies, including neurodegenerative diseases such as Parkinson's and Alzheimer's diseases. In this study, we used hydrogen/deuterium exchange, differential crosslinking, differential surface modification, and molecular dynamics (MD) simulations to characterize the structural changes in disordered proteins that result from ligand binding. We show here that both an ATP-independent protein chaperone, Spy L32P, and the FK506 binding domain of a prolyl isomerase, FKBP-25 F145A/I223P, are disordered, yet exhibit structures that are distinct from chemically denatured unfolded states in solution, and that they undergo transitions to a more structured state upon ligand binding. These systems may serve as models for the characterization of ligand-induced disorder-to-order transitions in proteins using structural proteomics approaches. SIGNIFICANCE: In this study, we used hydrogen/deuterium exchange, differential crosslinking, differential surface modification, and molecular-dynamics simulations to characterize the structural changes in disordered proteins that result from ligand binding. The protein-ligand systems studied here (the ATP-independent protein chaperone, Spy L32P, and the FK506 binding domain of a prolyl isomerase, FKBP-25 F145A/I223P) may serve as models for understanding ligand-induced disorder-to-order transitions in proteins. Additionally, the structural proteomic techniques demonstrated here are shown to be effective tools for the characterization of disorder-to-order transitions and can be used to facilitate study of other systems in which this class of structural transition can be used for modulating major pathological features of disease, such as the abnormal protein aggregation that occurs with Parkinson's disease and Alzheimer's disease.
对于无序蛋白质，配体结合可能是改变其结构动力学的关键事件。能够描述这种变化将有助于开发旨在稳定无序蛋白质的药物，这些蛋白质的错误折叠对于许多病理学很重要，包括神经退行性疾病，如帕金森病和阿尔茨海默病。在这项研究中，我们使用氢/氘交换、差异交联、差异表面修饰和分子动力学（MD）模拟来描述配体结合导致的无序蛋白质的结构变化。我们在这里表明，一种非 ATP 依赖性蛋白伴侣 Spy L32P 和一种脯氨酰异构酶 FKBP-25 F145A/I223P 的 FK506 结合结构域都是无序的，但它们的结构与溶液中化学变性的无规则状态明显不同，并且它们在配体结合后会发生向更结构化状态的转变。这些系统可作为使用结构蛋白质组学方法表征蛋白质中配体诱导的无序到有序转变的模型。意义：在这项研究中，我们使用氢/氘交换、差异交联、差异表面修饰和分子动力学模拟来描述配体结合导致的无序蛋白质的结构变化。这里研究的蛋白质-配体系统（非 ATP 依赖性蛋白伴侣 Spy L32P 和一种脯氨酰异构酶 FKBP-25 F145A/I223P 的 FK506 结合结构域）可作为理解蛋白质中配体诱导的无序到有序转变的模型。此外，这里展示的结构蛋白质组学技术被证明是描述无序到有序转变的有效工具，可用于促进对其他系统的研究，其中这种结构转变类可用于调节疾病的主要病理特征，如帕金森病和阿尔茨海默病中发生的异常蛋白聚集。
