Cryo-EM structure of a membrane-embedded glutamate receptor

膜嵌入谷氨酸受体的冷冻电镜结构

• 批准号: 8635183
• 负责人: FREDERICK J SIGWORTH
• 金额: $ 24.64万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8635183
• 关键词: Biochemistry; Computer software; Crystallization; Data Collection; Development; Environment; Excision; Freezing; Glutamate Receptor; Goals; Graph; Image; Imaging Techniques; Ion Channel; Laboratories; Life; Lipids; Liposomes; Membrane; Membrane Lipids; Membrane Potentials; Membrane Proteins; Methods; Modeling; Molecular Machines; Movement; Preparation; Probability; Process; Proteins; Research; Resolution; Roentgen Rays; Series; Signal Transduction; Specimen; Structure; Techniques; Vesicle; Work; X-Ray Crystallography; brain cell; computer program; electron crystallography; image processing; improved; interest; membrane model; method development; nanometer; particle; programs; public health relevance; reconstitution; reconstruction; structural biology; three dimensional structure; voltage

DESCRIPTION (provided by applicant): A long-pursued goal of structural biology has been the determination of the structure of membrane proteins in their native membrane environment, and in their various functional states as defined by transmembrane gradients of substrate molecules and membrane potentials. Application of such gradients, let alone crystallization in a membrane environment, has not been possible in the case of X-ray structure determination. The application of gradients has remained out of reach as well for the analysis of 2D crystals by electron crystallography. The proposed work concerns the further development of a general method for single-particle cryo-EM structure determination of membrane proteins reconstituted into liposomes. The method is called random spherically- constrained (RSC) single-particle reconstruction. In the proposed work we will work to surmount several remaining technical hurdles, and then apply this combination of biochemistry, EM imaging and image processing methods toward a nanometer- resolution structure of the GluA2 glutamate receptor as reconstituted into a lipid membrane.

描述（由申请人提供）：结构生物学长期追求的目标是确定膜蛋白在其天然膜环境中的结构，以及在由底物分子的跨膜梯度和膜电位定义的各种功能状态下的结构。在 X 射线结构测定的情况下，应用这种梯度是不可能的，更不用说在膜环境中结晶了。对于通过电子晶体学分析二维晶体而言，梯度的应用仍然遥不可及。拟议的工作涉及进一步开发用于重组脂质体的膜蛋白的单颗粒冷冻电镜结构测定的通用方法。该方法称为随机球面约束 (RSC) 单粒子重建。在拟议的工作中，我们将努力克服几个剩余的技术障碍，然后将生物化学、EM 成像和图像处理方法的组合应用于重构为脂质膜的 GluA2 谷氨酸受体的纳米分辨率结构。