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: 8723917
• 负责人: FREDERICK J SIGWORTH
• 金额: $ 20.6万
• 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/8723917
• 关键词: Bayesian Modeling; 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，以及由底物分子和膜电位的跨膜梯度定义的各种功能状态。这种梯度的应用，更不用说在膜环境中的结晶，在X射线结构测定的情况下是不可能的。梯度的应用对于通过电子晶体学分析2D晶体也仍然遥不可及。拟议的工作涉及进一步发展的一般方法，用于单颗粒冷冻-EM结构测定重组成脂质体的膜蛋白。该方法被称为随机球约束（RSC）单粒子重建。在所提出的工作中，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这种组合应用于重组成脂质膜的GluA 2谷氨酸受体的纳米分辨率结构。