XAS STUDIES OF METAL TRAFFICKING

这个子项目是利用由NIH/NCRR资助的中心拨款提供的资源的许多研究子项目之一。子项目和调查员(PI)可能从另一个NIH来源获得了主要资金，因此可能会出现在其他CRISE条目中。列出的机构是针对中心的，而不一定是针对调查员的机构。监测特定蛋白质对细胞事件的空间分布的能力使细胞生物学发生了革命性的变化，并使人们对疾病状态和蛋白质运输之间的关系有了更深的理解。共聚焦显微镜是推动蛋白质运输研究的技术，其基础是检测目标蛋白质的荧光标记抗体结合物的发射，使用激光激发来提供空间分辨率。目前的建议旨在应用替代光谱技术来监测蛋白质在细胞中的定位，使用蛋白质中的天然生色团来提供光谱信号。通常，这些发色团会与作为蛋白质基本催化成分的金属离子联系在一起。具体地说，我们的目标是开发空间分辨X射线吸收，作为金属蛋白活性部位结构的非侵入性探针，并将数据映射到更传统的蛋白质定位的共焦图像上。XAS能够提供活性中心金属离子化学形态的详细信息，这将为细胞中蛋白质和酶的成像提供更高水平的精确度。

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 ability to monitor the spatial distribution of select proteins in response to cellular events has revolutionized cell biology, and has led to a deeper understanding of the relationship between disease states and protein trafficking. Confocal microscopy, the technology that drives the study of protein trafficking, is based on detecting emission from fluorescently labeled antibody conjugates of the proteins of interest, using laser excitation to provide spatial resolution. The present proposal aims to apply alternative spectroscopic technologies to monitor the localization of proteins in cells, using native chromophores within the protein to provide the spectroscopic signal. Often these chromophores will be associated with metal ions which act as the essential catalytic component of the protein. Specifically our goal is to develop spatially resolved x-ray absorption as a non-invasive probe of metalloprotein active site structure, and to map the data onto more traditional confocal images of protein localization. The ability of XAS to provide details of the chemical speciation of the active site metal ion will provide a further level of sophistication to the imaging of proteins and enzymes in cells.