HIGH-THROUGHPUT X-RAY ABSORPTION SPECTROSCOPY FOR METALLOPROTEOMICS

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。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。列出的机构是 中心，不一定是研究者的机构。 我们建议与SSRL SMB光束线开发工作合作，提供一种称为高通量X射线吸收光谱（HTXAS）的新技术，旨在提供基于金属的蛋白质组视图。而功能基因组学（微阵列）技术询问蛋白质组成员的表达水平，HTXAS将提供有关金属蛋白质组成员的金属位点结构信息。这些技术分别解决了环境压力对某些蛋白质表达和蛋白质中某些金属位点组装的影响的生物学问题。作为这种发现科学方法的一个例子，我们建议将其应用于微生物生态学问题。特别是，微生物的金属蛋白质组补体在其生长环境中的重金属胁迫下如何变化？为此，我们正在使用激烈的热球菌作为模型，希瓦氏菌和耐金属罗尔斯通氏菌作为我们的第一个目标。所要求的射束时间用于该技术的可行性研究和与创建该技术相关的测试射束线开发活动。这项技术发展的副产品预计也将对“标准”生物XAS研究产生重大影响。

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.