XAS STUDIES OF METAL TRAFFICKING

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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.