Femtosecond nano-crystallography of membrane proteins

膜蛋白的飞秒纳米晶体学

• 批准号: 8818297
• 负责人: PETRA FROMME
• 金额: $ 28.9万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8818297
• 关键词: Biological; Cells; Code; Communities; Complex; Crystallography; Data; Data Analyses; Data Set; Detection; Development; Drug Targeting; Evaluation; Funding; Generations; Genes; Goals; Grant; Growth; Health; Human; Indium; Injection of therapeutic agent; Insecta; Investigation; Journals; Lasers; Legal patent; Life; Mediating; Medical; Membrane Proteins; Membrane Transport Proteins; Methods; Microfluidic Microchips; Microfluidics; Molecular; Nature; Paper; Pharmaceutical Preparations; Phase; Photosystem I; Physiologic pulse; Process; Proteins; Proteome; Publications; Publishing; Radiation; Reaction; Resolution; Roentgen Rays; Science; Shapes; Solid; Sorting - Cell Movement; Stream; Structural Biologist; Structure; Synchrotrons; Technology; Time; Work; X ray diffraction analysis; X-Ray Diffraction; base; cofactor; cytochrome c oxidase; design; direct application; free-electron laser; improved; in vivo; innovation; macromolecule; method development; movie; nano; nanocrystal; protein complex; protein structure; public health relevance; reconstruction; structural biology; technology development

DESCRIPTION (provided by applicant): This project aims to further develop the method of femtosecond nanocrystallography for the structure determination of membrane proteins. 30% of the human proteome consist of membrane proteins, which control and mediate the interaction between cells, regulate transport in and out of the cells and are also the major players in bioenergy conversion. Their importance for human health is overwhelming, with 60% of all current drugs being targeted to membrane proteins. Femtosecond crystallography is a new method for X-ray structure analysis of biological macromolecules, where hundreds of thousands of X-ray diffraction snapshots are collected from a stream of fully hydrated nano/microcrystals of proteins, using femtosecond X-ray pulses from a Free Electron Laser. The peak flux of an X-ray FEL is 109 times higher than the flux from a 3rd generation Synchrotron. While the X-ray pulses are so strong that they destroy any solid material, X-ray diffraction occurs before the biomolecules are destroyed. New developments in nanocrystal growth and characterization, together with development of new injector technology and data evaluation methods have progressed the new method of SFX at a very fast pace based on results from this project which has led to 16 publications, 9 of them in Nature of Science journals and one patent (pending). The proposal is focused on four major aims which include the development of new methods for nanocrystal growth and characterization (aim 1), innovations on new crystal sorting and Injector technology (aim 2), further development of data analysis methods including de novo phasing of SFX data (aim 3) and determination of membrane protein structures with SFX and new avenues for time-resolved SFX (aim 4). The goal is to open a new era in Structural Biology, where SFX is developed and used to solve challenging membrane protein structures. The development of time resolved SFX will provide new milestones towards the final goal to determine molecular movies of membrane proteins in action.

描述（由申请人提供）：该项目旨在进一步开发飞秒纳米晶体术的方法，以确定膜蛋白的结构。 30％的人蛋白质组由膜蛋白组成，这些膜蛋白控制和介导细胞之间的相互作用，调节流入和流入细胞，也是生物能转化的主要参与者。它们对人类健康的重要性是压倒性的，目前所有药物的目标均针对膜蛋白。飞秒晶体学是一种用于生物大分子的X射线结构分析的新方法，其中使用Femtosoctocond X射线脉冲从自由电子激光器中收集了数十万个X射线衍射快照，从完全水合的蛋白质/微晶体中收集了数十万个X射线衍射快照。 X射线FEL的峰值通量比第三代同步加速器的通量高109倍。尽管X射线脉冲非常强，以至于它们破坏了任何固体物质，但在生物分子被破坏之前发生X射线衍射。纳米晶生长和表征的新发展以及新的喷油器技术和数据评估方法的发展，基于该项目的结果，以非常快速的步伐发展了新的SFX方法，该项目导致了16个出版物，其中9种在科学期刊本质上和一项专利（待处理）。该提案的重点是四个主要目的，包括开发纳米晶增长和表征的新方法（AIM 1），有关新的水晶分类和喷油器技术的创新（AIM 2），进一步开发数据分析方法，包括从头逐渐相位的SFX数据（AIM 3）（AIM 3）（AIM 3）以及与SFX相关的膜蛋白结构以及SFX的确定以及SFX的确定，以sfx和新的verved sfx（Aim verved sfx（AIM）（AIM）（4）。目的是在结构生物学上开设一个新时代，在该时代开发SFX并用于解决挑战性的膜蛋白结构。时间解决的SFX的发展将为确定作用中膜蛋白的分子电影的最终目标提供新的里程碑。