HIGH-THROUGHPUT X-RAY ABSORPTION SPECTROSCOPY FOR METALLOPROTEOMICS

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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. We propose to partner with SSRL SMB beamline development efforts to provide a new technology, called high-throughput x-ray absorption spectroscopy (HTXAS), designed to provide a metal-based view of the proteome. Whereas functional genomics (microarray) technology interrogates the expression levels of the members of the proteome, HTXAS will provide metal-site structural information about members of the metalloproteome. These technologies address biological questions about the effect of environmental stress on expression of certain proteins and on assembly of certain metal sites in proteins, respectively. As one example of this discovery science approach, we propose to apply it to questions of microbial ecology. In particular, how does a microorganism's metalloproteome complement change under the stress of heavy metals in its growth environment? Toward that end, we are using Pyrococcus furiosus as a model and Shewanella oneidensis and Ralstonia metallidurans as our first targets. The requested beamtime is for feasibility studies of this technology and testing beamline development activities associated with creating this technology. Spinoffs of this technological development are expected to have a major impact on "standard" biological XAS studies also.