Structural Genomics on Membrane Proteins

该提案的总体目标是通过应用结构基因组学方法加速获取有关膜蛋白的结构信息，该方法由一组专家调查人员的集体经验提供信息。我们已经建立了纽约膜蛋白结构协会NYCOMPS，共同努力实现这一目标。NYCOMPS现在作为专业中心参与蛋白质结构倡议（PSI-2）的第2阶段。作为PSI-Biology的组成部分，NYCOMPS将由六个机构的12名主要研究者组成。我们的结构测定管道将通过对所有已知序列的生物信息学分析来选择靶点，然后进行重组DNA克隆，在细菌或真核细胞中表达蛋白质，并以中等高通量进行蛋白质纯化，然后继续通过X射线晶体学确定结构。我们位于纽约结构生物学中心（NYSBC）的蛋白质生产设施通过中等规模的纯化处理靶标;成功的候选物被分配给参与实验室进行放大和结晶。结构的功能分析将通过计算和常规实验生物化学进行。目标将通过提名从生物界，包括附属NYCOMPS成员，并从NYCOMPS生物主题，其中涉及阐明膜蛋白宇宙和结构研究能量稳态和代谢紊乱。一项技术开发计划将旨在提高管道效率和结果质量。该项目将被管理，以优化输出，并与PSI生物学网络和其他膜蛋白结构的努力有效地整合。

The overall objective of this proposal is to accelerate the acquisition of structural information about membrane proteins by applying a structural genomics approach informed by the collective experience of a team of expert investigators. We have established the New York Consortium on Membrane Protein Structure NYCOMPS to work together toward this objective. NYCOMPS participates as a Specialized Center in Phase 2 of the Protein Structure Initiative (PSI-2) now. As constituted for PSI-Biology, NYCOMPS will comprise 12 Principal Investigators at six institutions. Our pipeline for structure determination will select targets through a bioinformatics analysis of all known sequences, move on to recombinant DNA cloning, protein expression in bacteria or eukaryotic cells, and protein purification at moderately high throughput, and then continue on to determine structures by x-ray crystallography. Our Protein Production Facility at the New York Structural Biology Center (NYSBC) handles targets through purification at a mid-scale level; and successful candidates are distributed to participant laboratories for scale-up and crystallization. Functional analysis of structures will be performed both by computations and through routine experimental biochemistry. Targets will be identified through nominations from the biological community, including adjunct NYCOMPS members, and from NYCOMPS biological themes, which concern elucidation of the membrane protein universe and structural studies on energy homeostasis and metabolic disorders. A program in technology development will aim to improve pipeline efficiency and quality of results. The project will be managed to optimize output and to integrate effectively with the PSI-Biology network and with other membrane protein structure efforts.