HIGH-THROUGHPUT X-RAY ABSORPTION SPECTROSCOPY FOR METALLOPROTEOMICS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 我们建议与SSRL SMB光束线开发工作合作，提供一种名为高通量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。功能基因组学(微阵列)技术询问蛋白质组成员的表达水平，而HTXAS将提供关于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关于环境压力对某些蛋白质表达的影响和对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重金属胁迫下如何在其生长环境中发生变化？为此，我们以狂热杆菌为模型，以单氏希瓦氏菌和金属拉尔斯氏菌为第一目标。申请的波束时间用于这项技术的可行性研究和测试与创造这项技术相关的波束线开发活动。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.