XAS STUDIES OF METAL TRAFFICKING

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，不一定是研究者所在机构。监测响应细胞事件的选择蛋白质的空间分布的能力已经彻底改变了细胞生物学，并且已经导致对疾病状态和蛋白质运输之间的关系的更深入理解。共聚焦显微镜是一种推动蛋白质运输研究的技术，它基于检测感兴趣蛋白质的荧光标记抗体缀合物的发射，使用激光激发来提供空间分辨率。本提案旨在应用替代的光谱技术来监测细胞中蛋白质的定位，使用蛋白质内的天然发色团来提供光谱信号。通常这些发色团将与金属离子结合，金属离子作为蛋白质的基本催化组分。具体来说，我们的目标是开发空间分辨的X射线吸收作为金属蛋白活性位点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聚焦图像上。XAS提供活性位点金属离子的化学形态的详细信息的能力将为细胞中蛋白质和酶的成像提供更高水平的复杂性。

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.