STRUCTURAL AND FUNCTIONAL GENOMICS OF PROTEIN FAMILIES

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。 本项目中在PXRR内开展的数据收集活动侧重于中等吞吐量结构的确定和表征，主要是原核生物和真核生物来源的蛋白质的酶机制，这些蛋白质参与了各种生物过程。这些项目的具体目标是：(A)确定脱脂蛋白的结构；(B)确定与各种可能的底物/产物/抑制物或辅因子分子的共晶结构，以阐明酶的机制；以及(C)对这些蛋白质进行额外的生物物理和生化表征以及定点突变，以便更好地了解它们的功能和补充晶体结构。 已知和未知功能的细菌蛋白主要选自三个已测序的大肠杆菌基因组。其中一些蛋白质是大肠杆菌致病菌株以及其他致病细菌所特有的，可能是治疗干预的潜在靶点。从这些研究中产生了30多种大肠杆菌蛋白质的晶体结构，代表了广泛的代谢过程。除了大肠杆菌蛋白，我们还对幽门螺杆菌和空肠弯曲菌参与假氨基酸等碳水化合物合成的细菌酶和一般的N-糖基化机制感兴趣。最后，我们对肝素黄杆菌、类杆菌和其他物种的糖胺聚糖裂解酶有着长期的兴趣。我们发表的大多数结果都是基于一种天然酶的结构和几个复合体。 真核蛋白质是根据蒙特利尔蛋白质组学网络(如CREG，EnthoProtein的第10结构域)携带的细胞器的蛋白质组学分析以及大型NRCC项目携带的正常和癌细胞系的比较微阵列数据来选择的。在用转化生长因子-b治疗作为转移模型的细胞系之后，选择了感兴趣的靶点。选择显著上调或下调的基因进行结构研究。此外，我们对信号通路中的支架蛋白及其与结合伙伴的复合体(例如MP1-p14复合体)感兴趣。

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 data collection activities carried out within the PXRR in this project are focused on medium-throughput structure determination and characterization, primarily of enzymatic mechanisms of proteins from prokaryotic and eukaryotic sources that are involved in a variety of biological processes. The specific goals of these projects are to (a) determine the structure of the apo-protein (b) determine the co-crystal structure with a variety of possible substrate/product/inhibitor or cofactor molecules to clarify the enzymatic mechanism and (c) perform additional biophysical and biochemical characterization and site-directed mutagenesis of these proteins in order to better understand their function and to complement the crystal structure. Bacterial proteins of both known and unknown function are predominantly selected from the three sequenced genomes of Escherichia coli. Some of the proteins are unique to both E. coli pathogenic strains, as well as to other pathogenic bacteria, and may represent potential targets for therapeutic intervention. Over 30 crystal structures of E. coli proteins have arisen from this these studies, and represent a broad range of metabolic processes. In addition to E. coli proteins, we are interested in bacterial enzymes from Helicobacter pylori and Campylobacter jejuni involved in pseudaminic acid other carbohydrates synthesis and the general N-glycosylation machinery. Finally, we have a long standing interest in glycosaminoglycan lyases from Flavobacterium heparinum, Bacteroides thetaiotaomicron and other species Most of our published results are based on structure of a native enzyme together with several complexes. Eukaryotic proteins are selected based on proteomics analysis of cellular organelles carried within the Montreal Proteomics Network (e.g. CREG, ENTH domain of enthoprotin) and based on comparative microarray data for normal and cancer cell lines carried within a large-scale NRCC project. Targets of interest were selected following TGF-b treatment of a cell line used as a model for metastasis. Genes that are signifcantly up- or down-regulated were selected for structural studies. In addition, we are interested in scaffolding proteins in signaling pathways and their complexes with binding partners (e.g. MP1-p14 complex).