Membrane Proteins. Crystallogenesis and X-ray Structure

膜蛋白。

• 批准号: 7683370
• 负责人: MARTIN D. CAFFREY
• 金额: $ 1.5万
• 依托单位: UNIVERSITY OF LIMERICK
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-23 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7683370
• 关键词: 3-Dimensional; Adenosine; Archives; Arts; Bacteriorhodopsins; Behavior; Benchmarking; Biology; Bos taurus; Cattle; Cell physiology; Characteristics; Chemicals; Circular Dichroism; Code; Colicin E3; Collaborations; Complex; Crystal Formation; Crystallization; Crystallography; Cystic Fibrosis; Cytochrome P450; Data; Databases; Disease; Environment; Escherichia coli; Eukaryota; Eukaryotic Cell; Family; Figs - dietary; Focus Groups; G-Protein-Coupled Receptors; GLUT-1 protein; Growth; Harvest; Health; Homologous Gene; Housing; Human; Human Genome; Hydration status; Hydroquinones; Inclusion Bodies; Individual; Ions; Knowledge; Light; Lipid Bilayers; Lipids; Maps; Measures; Membrane; Membrane Proteins; Metals; Methods; Molecular; Molecular Structure; Monoglycerides; NMR Spectroscopy; Neutrons; Nitrite Reductase; Numbers; Organic Chemistry; Oxidases; Peptides; Performance; Peripheral; Phase; Process; Prokaryotic Cells; Proteins; Proteomics; Protocols documentation; Range; Reaction; Replication Initiation; Research Personnel; Residual state; Resolution; Rhodopsin; Robot; Robotics; Roentgen Rays; Role; Role playing therapy; Signal Transduction; Staging; Structure; Surface; Synchrotrons; System; Testing; Thermus thermophilus; Transducin; United States National Institutes of Health; Vitamin B 12; Work; base; cytochrome c oxidase; cytochrome caa(3); design; improved; insight; interest; method development; nitric oxide reductase; novel; programs; protein folding; protein function; protein structure; quinol fumarate reductase; receptor; reconstitution; signal peptidase; structural biology; success; trafficking

DESCRIPTION (provided by applicant): Solving the structure with a view to understanding the function of membrane proteins remains one of the grand challenges in all of Biology. Given that a third of the human genome codes for membrane proteins, many of which serve signal transducing, structural and transport roles and are targets of disease causative and treatment agents, the health consequences of meeting the challenge are great. A multifaceted approach will be taken to advance our understanding of membrane protein function by producing structure grade crystals for use in macromolecular crystallography. Emphasis is placed on crystallization in lipidic mesophases by what is referred to as the 'in meso' or cubic phase method. This is proving to be a generally useful approach for membrane protein structure determination. With this method crystallization takes place in a membrane environment that is likely to be favored by the target membrane protein. We have built a unique, state-of-the-art robot that performs in meso crystallization in high-throughput fashion and that requires miniscule amounts of protein, lipid and precipitant. It will be used in the current application to produce 3-D crystals for the high-resolution structure determination of a select group of membrane proteins. The target group covers a broad range of membrane protein types including eukaryote and prokaryote, integral and peripheral, monomeric and multimeric, as well as protein-protein and protein-peptide complexes. Some of the target proteins will be produced in-house, some will be provided by individual collaborators, while others will be supplied through the NIH Structural Proteomics Initiative. In line with the NIH Structural Biology Road Map, and in parallel with the proposed structure study, effort will be devoted to improving crystallogenesis and to broadening the range of membrane protein targets that yield to structure determination. This will be achieved by implementing a synthesis program whereby lipids with desirable physico-chemical characteristics are produced for use in crystallization trials. The molecular mechanism of crystal nucleation and growth will be studied too with a view to more rational and successful crystallogenesis. Success in obtaining crystals, and ultimately the atomic structure of membrane proteins, will enhance our understanding of some of the most fundamental processes underlying cellular function that are integral to human health.

描述（由申请人提供）：解决结构以了解膜蛋白的功能仍然是整个生物学的重大挑战之一。鉴于人类基因组的三分之一编码膜蛋白，其中许多膜蛋白发挥信号转导、结构和运输作用，并且是致病和治疗药物的靶标，应对这一挑战对健康的影响是巨大的。将采取多方面的方法，通过生产用于大分子晶体学的结构级晶体来增进我们对膜蛋白功能的理解。重点是通过所谓的“中间相”或立方相方法在脂质中间相中结晶。事实证明，这是一种普遍有用的膜蛋白结构测定方法。通过这种方法，结晶发生在可能受到目标膜蛋白青睐的膜环境中。我们建造了一种独特的、最先进的机器人，它以高通量方式进行介观结晶，并且需要极少量的蛋白质、脂质和沉淀剂。它将在当前的应用中用于生产 3D 晶体，以用于选定一组膜蛋白的高分辨率结构测定。目标群体涵盖广泛的膜蛋白类型，包括真核生物和原核生物、整体和外周、单体和多聚体，以及蛋白质-蛋白质和蛋白质-肽复合物。一些目标蛋白将在内部生产，一些将由个人合作者提供，而另一些将通过 NIH 结构蛋白质组计划提供。 根据 NIH 结构生物学路线图，并与拟议的结构研究并行，将致力于改善晶体发生并扩大可进行结构测定的膜蛋白靶标范围。这将通过实施合成计划来实现，通过该计划生产具有所需物理化学特性的脂质用于结晶试验。晶体成核和生长的分子机制也将得到研究，以期更加合理和成功的晶体发生。成功获得晶体并最终获得膜蛋白的原子结构，将增强我们对细胞功能背后的一些最基本过程的理解，这些细胞功能对人类健康至关重要。