Structural Genomics and Membrane Proteins

本提案的总体目标是加快获取以下结构信息： 膜蛋白通过应用结构基因组学方法，由团队的集体经验告知 专家调查员。我们建立了纽约膜蛋白结构协会 ��NYCOMPS为实现这一目标而共同努力。NYCOMPS作为专业中心参加了 蛋白质结构倡议PSI 2作为构成PSI生物学，NYCOMPS将包括12个主要 六个机构的调查员。我们的结构确定管道将通过 对所有已知序列进行生物信息学分析，然后进行重组DNA克隆、蛋白质表达 细菌或真核细胞，并以中等高通量进行蛋白质纯化，然后继续进行 用X射线晶体学确定结构。我们位于纽约结构性建筑的蛋白质生产设施 生物中心NYSBC通过中等规模水平的纯化处理靶标;成功的候选者是 分发给参与实验室进行规模放大和结晶。结构的功能分析将 通过计算和常规的实验生物化学来完成。将确定目标 通过生物界的提名，包括NYCOMPS的附属成员，以及NYCOMPS的提名， 生物学主题，涉及膜蛋白宇宙的阐明和能量的结构研究 体内平衡和代谢紊乱。一项技术开发计划将旨在改善管道 效率和成果质量。将对该项目进行管理，以优化产出，并有效地与 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 perfomed 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.