Femtosecond nano-crystallography of membrane proteins

膜蛋白的飞秒纳米晶体学

基本信息

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
项目类别：
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

项目摘要

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（申请人提供）：该项目旨在进一步开发用于膜蛋白结构测定的飞秒纳米晶体学方法。人类蛋白质组的 30% 由膜蛋白组成，膜蛋白控制和介导细胞之间的相互作用，调节进出细胞的运输，也是生物能量转换的主要参与者。它们对人类健康的重要性是压倒性的，目前所有药物中 60% 都是针对膜蛋白的。飞秒晶体学是一种用于生物大分子 X 射线结构分析的新方法，其中使用来自自由电子的飞秒 X 射线脉冲从完全水合的蛋白质纳米/微晶体流中收集数十万个 X 射线衍射快照激光。 X 射线 FEL 的峰值通量比第三代同步加速器的通量高 109 倍。虽然 X 射线脉冲非常强，可以破坏任何固体材料，但 X 射线衍射会在生物分子被破坏之前发生。纳米晶体生长和表征方面的新进展，加上新注射器技术和数据评估方法的发展，基于该项目的结果，以非常快的速度发展了 SFX 新方法，该项目已发表 16 篇论文，其中 9 篇发表在《自然》杂志上科学期刊和一项专利（正在申请中）。该提案重点关注四个主要目标，包括开发纳米晶体生长和表征的新方法（目标 1）、新晶体分选和注射器技术的创新（目标 2）、进一步开发数据分析方法，包括 SFX 的从头定相数据（目标 3）并使用 SFX 确定膜蛋白结构以及时间分辨 SFX 的新途径（目标 4）。其目标是开启结构生物学的新时代，开发 SFX 并用于解决具有挑战性的膜蛋白结构。时间分辨 SFX 的开发将为确定膜蛋白起作用的分子电影的最终目标提供新的里程碑。