Department of Biochemistry and Pharmacology, Bio21 Molecular Science and Biotechnology Institute, The University of Melbourne, Parkville, VIC, 3010, Australia.
Department of Chemistry, University of Cambridge, Cambridge, CB2 1EW, United Kingdom.
NPJ Syst Biol Appl. 2022 Nov 28;8(1):46. doi: 10.1038/s41540-022-00256-3.
The correct spatio-temporal organization of the proteome is essential for cellular homeostasis. However, a detailed mechanistic understanding of this organization and how it is altered in response to external stimuli in the intact cellular environment is as-yet unrealized. 'Protein painting methods provide a means to address this gap in knowledge by monitoring the conformational status of proteins within cells at the proteome-wide scale. Here, we demonstrate the ability of a protein painting method employing tetraphenylethene maleimide (TPE-MI) to reveal proteome network remodeling in whole cells in response to a cohort of commonly used pharmacological stimuli of varying specificity. We report specific, albeit heterogeneous, responses to individual stimuli that coalesce on a conserved set of core cellular machineries. This work expands our understanding of proteome conformational remodeling in response to cellular stimuli, and provides a blueprint for assessing how these conformational changes may contribute to disorders characterized by proteostasis imbalance.
蛋白质组的正确时空组织对于细胞内稳态至关重要。然而，对于这种组织的详细机制理解以及它如何在完整的细胞环境中响应外部刺激而改变，目前尚未实现。“蛋白质绘图方法通过在蛋白质组范围内监测细胞内蛋白质的构象状态，为解决这一知识空白提供了一种手段。在这里，我们展示了一种使用四苯乙烯马来酰亚胺 (TPE-MI) 的蛋白质绘图方法的能力，该方法能够揭示整个细胞中蛋白质组网络重塑对一系列具有不同特异性的常用药理学刺激的反应。我们报告了对单个刺激的特定但异质的反应，这些反应集中在一组保守的核心细胞机制上。这项工作扩展了我们对蛋白质组在响应细胞刺激时构象重塑的理解，并为评估这些构象变化如何有助于以蛋白质稳态失衡为特征的疾病提供了蓝图。
