IN-SITU UV/VIS SPECTROSCOPY FOR MACROMOLECULAR CRYSTALLOGRAPHY

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定为研究者机构。该项目的目标是能够同时获取X射线衍射数据和紫外/可见光和红外区域的单晶吸收光谱。在UV（190 - 400 nm）和可见光（400 - 800 nm）区域的吸收光谱的分析提供了关于蛋白质在结晶状态时的电子状态的信息。在衍射实验期间，将分析光谱测量以确认氧化还原状态或反应中间体的存在，或潜在地监测可能通过光解或X射线产生的降解产物。在BL 9 -2上设置单晶显微分光光度计，以记录测角仪上晶体的光谱。光学器件和第二显微镜的定位经过改进以便于使用。 从几个晶体系统中记录光谱的尝试为开发过程提供了重要的反馈，特别是定义如何改进定位系统，为光学系统增加额外的自由度，以实现光束的精确对准和聚焦。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of this project is to enable the simultaneous acquisition of x-ray diffraction data and single crystal absorption spectra in the UV/Visible and infrared regions. Analysis of the absorption spectrum in the UV (190 - 400 nm) and visible (400 - 800 nm) regions provides information about the electronic state of proteins while in the crystalline state. During diffraction experiments, spectroscopic measurements will be analyzed to confirm the redox state or the presence of reaction intermediates or to potentially monitor degradation products that may be produced either by photolysis or by x-rays. A single crystal microspectrophotometer was set up on BL9-2 to record spectra from crystals on the goniometer. Positioning of the optics and a 2nd microscope were refined for ease of use. Attempts at recording spectra from several crystal systems provided important feedback for the development process, in particular to define how the positioning system will be refined, adding additional degrees of freedom to the optics for precise alignment and focusing of the light beam.