The structure-function relationships of biomolecules have captured the interest and imagination of the scientific community and general public since the field of structural biology emerged to enable the molecular understanding of life processes. Proteins that play numerous functional roles in cellular processes have remained in the forefront of research, inspiring new characterization techniques. In this review, we present key theoretical concepts and recent experimental strategies using femtosecond stimulated Raman spectroscopy (FSRS) to map the structural dynamics of proteins, highlighting the flexible chromophores on ultrafast timescales. In particular, wavelength-tunable FSRS exploits dynamic resonance conditions to track transient-species-dependent vibrational motions, enabling rational design to alter functions. Various ways of capturing excited-state chromophore structural snapshots in the time and/or frequency domains are discussed. Continuous development of experimental methodologies, synergistic correlation with theoretical modeling, and the expansion to other nonequilibrium, photoswitchable, and controllable protein systems will greatly advance the chemical, physical, and biological sciences.

中文翻译：

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飞秒激发拉曼光谱法绘制蛋白质的结构动力学图。

自从结构生物学领域出现以使人们能够对生命过程进行分子理解以来，生物分子的结构-功能关系就引起了科学界和公众的兴趣和想象。在细胞过程中发挥许多功能作用的蛋白质一直处于研究的前沿，激发了新的表征技术。在这篇综述中，我们介绍了使用飞秒激发拉曼光谱（FSRS）绘制蛋白质结构动力学的关键理论概念和最新实验策略，突出了超快时间尺度上的柔性发色团。尤其是，波长可调FSRS利用动态共振条件来跟踪瞬态物种相关的振动运动，从而进行合理的设计来改变功能。讨论了在时域和/或频域中捕获激发态生色团结构快照的各种方法。实验方法的不断发展，与理论模型的协同相关以及向其他非平衡，光开关和可控蛋白质系统的扩展将极大地推动化学，物理和生物学科学的发展。