EAGER: Methodology development for 3D atomic-scale structural dynamics movies of enzymes

EAGER：酶的 3D 原子尺度结构动力学电影的方法开发

基本信息

批准号：
1842951
负责人：
Shimon Weiss
金额：
$ 30万
依托单位：
University of California-Los Angeles
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-02-28
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=1842951&HistoricalAwards=false
关键词：
EAGER Methodology development 3D atomic

项目摘要

Capturing the dynamic structure of a biological machine performing its function remains the holy grail of biology. Conventional tools provide only structural "snapshots" of stable states along the reaction pathway. This project aims to develop methods that will capture a full movie of a macromolecule's structure as it works. The research tools developed will be applicable to a wide variety of biological systems, and will be disseminated by means of publications, tutorials, conference presentations and the like, to accelerate their adoption. The education component of the project will emphasize the use of single molecule biophysics and computational tools for dynamic structure determination. It will provide high school, undergraduate, graduate students and postdocs training in state-of-the-art single molecule biophysics research, and offer leadership experiences for graduate students and postdocs as well.Specifically, this project will study the structure of the transcription bubble in a simple bacterial transcription system during initiation as a test case for developing new technological concepts. The existence of certain intermediates in bacterial transcription suggests common features of the process exist across the different domains of life and reflects a conserved regulatory mechanism; thus, the findings will have broad utility. For the purpose of dynamic structure determination, the project will combine experimental and theoretical tools, including multi-spot single-molecule FRET measurements, fast single-molecule microfluidic mixing, molecular dynamics simulations and time-resolved single-photon detection to acquire data on- and perform molecular modeling of- the transcription bubble during initiation.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

捕获执行其功能的生物机器的动态结构仍然是生物学的圣杯。传统工具仅提供反应路径中稳定状态的结构“快照”。该项目旨在开发能够捕捉大分子结构工作过程的完整影片的方法。开发的研究工具将适用于各种生物系统，并将通过出版物、教程、会议演讲等方式传播，以加速其采用。该项目的教育部分将强调使用单分子生物物理学和计算工具来确定动态结构。它将为高中生、本科生、研究生和博士后提供最先进的单分子生物物理学研究培训，并为研究生和博士后提供领导经验。具体来说，该项目将研究转录气泡的结构在启动过程中使用简单的细菌转录系统作为开发新技术概念的测试用例。细菌转录中某些中间产物的存在表明该过程在生命的不同领域存在共同特征，并反映了保守的调节机制；因此，研究结果将具有广泛的用途。为了确定动态结构，该项目将结合实验和理论工具，包括多点单分子 FRET 测量、快速单分子微流体混合、分子动力学模拟和时间分辨单光子检测，以获取有关数据该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。