Caramello Nicolas, Adam Virgile, Pearson Arwen R, Royant Antoine
European Synchrotron Radiation Facility 71 Avenue des Martyrs CS 40220 38043Grenoble Cedex 9 France.
Institute for Nanostructure and Solid State Physics Universität Hamburg, Center for Ultrafast Imaging HARBOR, Bldg 610, Luruper Chaussee 149 Hamburg22761 Germany.
J Appl Crystallogr. 2025 May 31;58(Pt 3):1068-1078. doi: 10.1107/S1600576725003541. eCollection 2025 Jun 1.
Over the past ten years, there has been a surge in the demand for optical spectroscopy (OS), since optical spectroscopy is one of the few biophysical characterization methods applicable to both protein solutions and crystals. Historically, OS has been used to compare the state of proteins in crystals and in solution, and to assess their functionality by determining the redox state of metal ions, cofactors or chromophores. The recent rejuvenation of time-resolved crystallography experiments has sparked a renewed interest in optical spectroscopy as a bridge between kinetic studies in solution and in the crystalline state. The method of OS can be defined as the ensemble of spectroscopic techniques in the UV-visible-infrared range that can be applied to crystals. It has also been instrumental in understanding specific X-ray radiation damage to redox-sensitive parts of proteins. Spectra recorded from crystals are affected by crystal orientation, shape or position due to various optical phenomena. Fortunately, these can be modelled and their effect can be corrected. The OS laboratory at the European Synchrotron Radiation Facility (ESRF) specializes in recording UV-Vis absorption, fluorescence emission and Raman spectra from protein crystals. Here, we present a suite of utilities that streamline the analysis and correction of UV-Vis absorption OS data, encased in a graphical interface. This was originally developed for the OS laboratory at ESRF but is available as a standalone package, with the aim of making OS more accessible.
在过去十年中，对光学光谱学（OS）的需求激增，因为光学光谱学是少数适用于蛋白质溶液和晶体的生物物理表征方法之一。从历史上看，OS已被用于比较晶体和溶液中蛋白质的状态，并通过确定金属离子、辅因子或发色团的氧化还原状态来评估其功能。时间分辨晶体学实验最近的复兴引发了人们对光学光谱学的新兴趣，它作为溶液和晶体状态动力学研究之间的桥梁。OS方法可以定义为可应用于晶体的紫外-可见-红外范围内的光谱技术的集合。它在理解蛋白质氧化还原敏感部分的特定X射线辐射损伤方面也发挥了作用。由于各种光学现象，从晶体记录的光谱会受到晶体取向、形状或位置的影响。幸运的是，这些可以被建模，其影响也可以被校正。欧洲同步辐射装置（ESRF）的OS实验室专门记录蛋白质晶体的紫外-可见吸收、荧光发射和拉曼光谱。在这里，我们展示了一套实用程序，这些程序简化了紫外-可见吸收OS数据的分析和校正，并封装在一个图形界面中。它最初是为ESRF的OS实验室开发的，但现在作为一个独立的软件包提供，目的是使OS更容易使用。
