Ruan Hao, Lemke Edward A
BioCenter, Johannes Gutenberg University Mainz, Mainz, Germany; email:
IMB Postdoc Program, Institute of Molecular Biology, Mainz, Germany.
Annu Rev Phys Chem. 2025 Apr;76(1):103-128. doi: 10.1146/annurev-physchem-082423-030632.
Investigating protein dynamic structural changes is fundamental for understanding protein function, drug discovery, and disease mechanisms. Traditional studies of protein dynamics often rely on investigations of purified systems, which fail to capture the complexity of the cellular environment. The intracellular milieu imposes distinct physicochemical constraints that affect macromolecular interactions and dynamics in ways not easily replicated in isolated experimental setups. We discuss the use of fluorescence resonance energy transfer, fluorescence anisotropy, and minimal photon flux imaging technologies to address these challenges and directly investigate protein conformational dynamics in mammalian cells. Key findings from the application of these techniques demonstrate their potential to reveal intricate details of protein conformational plasticity. By overcoming the limitations of traditional in vitro methods, these approaches offer a more accurate and comprehensive understanding of protein function and behavior within the complex environment of mammalian cells.
研究蛋白质动态结构变化是理解蛋白质功能、药物发现和疾病机制的基础。传统的蛋白质动力学研究通常依赖于对纯化系统的研究，而这种研究无法捕捉细胞环境的复杂性。细胞内环境施加了独特的物理化学限制，以不易在孤立实验装置中复制的方式影响大分子相互作用和动力学。我们讨论了使用荧光共振能量转移、荧光各向异性和最小光子通量成像技术来应对这些挑战，并直接研究哺乳动物细胞中的蛋白质构象动力学。这些技术应用的关键发现证明了它们揭示蛋白质构象可塑性复杂细节的潜力。通过克服传统体外方法的局限性这些方法能更准确、全面地理解蛋白质在哺乳动物细胞复杂环境中的功能和行为。
