CHARACTERIZATION OF LIPID CUBIC PHASE MATRICES FOR MEMBRANE PROTEIN CRYSTALLIZAT

膜蛋白结晶脂质立方相基质的表征

• 批准号: 8170236
• 负责人: HIROTSUGU TSURUTA
• 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/8170236
• 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; Lipids; Measures; Membrane Proteins; Phase; Preparation; Property; Proteins; Research; Research Personnel; Resolution; Resources; Sampling; Source; Techniques; United States National Institutes of Health; novel

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. 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 SSRL BL4-2. 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.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 膜蛋白的结构研究受到衍射型高分辨率晶体可用性的限制。在脂质立方相(LCP)基质中(或在介观中)结晶已被证明可以产生高质量的具有挑战性的膜蛋白晶体，如人G蛋白偶联受体。介观技术的广泛应用要求鉴定具有特定相性质的新脂类，能够稳定具有大范围大小和结构的蛋白质。在用于特定应用之前，应对新型脂质基质的相和结构行为进行彻底的表征。我们建议通过在96孔夹心板中制备样品并在SSRLBL4-2上进行原位测量来克服与用于X射线研究的传统脂质样品制备相关的障碍。这种方法将使我们能够在模拟结晶试验中遇到的条件下，以高通量模式筛选洗涤剂、添加剂脂类、蛋白质以及各种沉淀剂对脂质基质的影响。