Structural Genomics on Membrane Proteins

本提案的总体目标是通过应用结构基因组学方法来加速获得膜蛋白的结构信息，该方法由一组专家研究人员的集体经验提供信息。为了实现这一目标，我们已经建立了纽约膜蛋白结构协会（NYCOMPS）。NYCOMPS现在作为一个专业中心参与蛋白质结构倡议（PSI-2）的第二阶段。NYCOMPS是为psi -生物学设立的，将由6个机构的12名首席研究员组成。我们的结构测定流水线将通过对所有已知序列的生物信息学分析来选择目标，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。我们在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目标；成功的候选人将被分配到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的生物化学实验来进行。靶点将通过生物界的提名来确定，包括NYCOMPS的附属成员，以及NYCOMPS的生物学主题，这些主题涉及膜蛋白宇宙的阐明以及能量稳态和代谢紊乱的结构研究。一项技术开发计划旨在提高管道效率和结果质量。该项目将设法优化输出，并有效地整合psi -生物学网络和其他膜蛋白结构的努力。

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.