TEMPERATURE-DEPENDENT CONFORMATIONAL SAMPLING OF NATIVELY UNFOLDED NUCLEOPORINS

天然解折叠核孔蛋白的温度依赖性构象采样

• 批准号: 8170314
• 负责人: MICHAEL F REXACH
• 金额: $ 0.1万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170314
• 关键词: Architecture; Behavior; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Detergents; Funding; G-Protein-Coupled Receptors; Grant; Human; In Situ; Institution; Joints; Lipids; Measures; Membrane Proteins; Nuclear Pore Complex Proteins; Phase; Preparation; Property; Proteins; Research; Research Personnel; Resolution; Resources; Sampling; Source; Techniques; Technology; Temperature; United States National Institutes of Health; Work; beamline; design; innovation; novel; research study

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. Structural studies of membrane proteins are limited by the availability of crystals diffracting to high resolution. Crystallization in lipidic cubic phase (LCP) matrices (or in meso) has proven to yield high quality crystals of challenging membrane proteins, such as human G protein-coupled receptors. Broader applications of in meso techniques require identification of new lipids with specific phase properties capable of stabilizing proteins with large range of sizes and architectures. At the Joint Center for Innovative Membrane Protein Technologies (JCIMPT) we are working on design and synthesis of such lipids. The phase and structural behavior of novel lipid matrices should be thoroughly characterized prior to being used in specific applications. We propose to overcome the obstacles associated with conventional preparation of lipid samples for x-ray studies, by preparing samples in 96-well sandwich plates and measuring them in situ at the BioSAXS beamline. This approach will allow us to screen for effects of detergents, additive lipids, proteins as well as great variety of precipitants on the lipidic matrices in the high-throughput mode at conditions mimicking those encountered during crystallization trials. During the duration of this proposal we anticipate to fully characterize 10-12 most promising novel lipids pre-selected out of a larger pool of synthesized candidates. The obtained results will be indispensable for selection of proper lipids and guiding in meso crystallization experiments.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 膜蛋白的结构研究受到衍射到高分辨率的可用性的限制。事实证明，脂质立方相（LCP）矩阵（或中）的结晶已被证明可以产生具有挑战性膜蛋白（例如人G蛋白偶联受体）的高质量晶体。在中索技术中的更广泛应用需要鉴定具有能够稳定具有较大尺寸和架构的蛋白质的特定相特性的新脂质。在创新膜蛋白技术联合中心（JCIMPT），我们正在研究此类脂质的设计和合成。在特定应用中使用之前，应彻底表征新型脂质矩阵的相位和结构行为。 我们建议通过在96孔三明治板上制备样品并在Biosaxss束线上进行测量，以克服与脂质样品进行常规制备进行X射线研究相关的障碍。 这种方法将使我们能够在高通量模式下在模仿结晶试验期间遇到的那些情况下，在高通量模式下，在高通量模式下，在脂质矩阵上筛选了脂质矩阵上的洗涤剂，添加剂脂质，蛋白质以及各种降水剂的影响。在该提案的持续时间内，我们预计将充分表征10-12个最有前途的新型脂质，从较大的合成候选物中预先选择的脂质。获得的结果对于选择适当的脂质并在中索结晶实验中是必不可少的。