Triggered functional dynamics of proteins in biomimetic environments by time-resolved electron paramagnetic resonance at very high magnetic fields

通过极高磁场下的时间分辨电子顺磁共振触发仿生环境中蛋白质的功能动力学

• 批准号: 2025860
• 负责人: Mark Sherwin
• 金额: $ 75万
• 依托单位: University of California-Santa Barbara
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2025860&HistoricalAwards=false
• 关键词: Triggered; functional; dynamics; proteins; biomimetic

Proteins are the nano-scopic molecular machines that power life as we know it. Thanks to a revolution in structural biology, we now know the three-dimensional shapes of more than 150,000 proteins. However, to fully understand the operation of proteins as machines, one must understand how they move after being triggered by an external event such as attaching to a particular molecule, movement of a neighboring protein, or a change in voltage or light intensity. A particularly exciting class of proteins called photo responsive proteins can move to control the behaviors of cells when exposed to light. This research will develop an advanced form of magnetic resonance—of which the most familiar application is magnetic resonance imaging (MRI)—to make “movies” of the motion of photo responsive proteins in response to a flash of light. The ability to “film” protein motion will provide critical tools to guide widespread efforts to engineer and optimize proteins for important applications ranging from sustainable manufacturing to controlling nerves with light. This project will engage next generation of scientists by public outreach activities through the “Questboard” developed by the PI’s group. Proteins are the molecular machines that power life as we know it. The rapidly-growing protein data bank now holds more than 150,000 protein structures. However, to fully understand the operation of proteins as machines, one must understand their triggered functional dynamics--how protein structures evolve in time after being triggered by an external event such as ligand docking, movement of a neighboring protein, or a change in voltage or light intensity. Photo responsive proteins are a particularly exciting class of proteins that, after absorbing a photon, generate mechanical energy to modulate biochemical processes and cellular behavior. This project will support convergent research to measure the time-resolved conformational changes triggered by a pulse of light in two photo responsive proteins: a microbial rhodopsin proton pump, and optogenetic proteins called Light, Oxygen, and Voltage (LOV) sensing domains. The measurements are enabled by a unique methodology, under development over the last decade by the PI and co-PI, which combines electron paramagnetic resonance (EPR) at very high magnetic fields (8.6 Tesla) and frequencies (240 GHz) with site-directed mutagenesis and spin labeling using compounds based on Gd (III) paramagnetic metal centers. This project is supported by the Molecular Biophysics Cluster of the Molecular and Cellular Biosciences Division in the Biological Sciences Directorate.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.

蛋白质是我们所知的纳米震源分子机器。感谢您在结构生物学的革命中，我们现在知道超过150,000种蛋白质的三维形状。但是，要充分理解蛋白质作为机器的操作，必须了解它们在被外部事件触发后如何移动，例如附着在特定分子上，相邻蛋白质的运动或电压或光强度的变化。一类特别令人兴奋的蛋白质称为照片反应蛋白可以在暴露于光线时移动以控制细胞的行为。这项研究将开发出一种先进的磁共振形式（最熟悉的应用是磁共振成像（MRI）），以使照片反应式蛋白质的运动“电影”以响应光闪光。 “胶卷”蛋白质运动的能力将提供关键的工具，以指导宽度努力，以设计和优化蛋白质，以适应从可持续制造到光线控制神经的重要应用。该项目将通过PI小组开发的“ Questboard”通过公共外展活动与下一代科学家联系。蛋白质是我们所知道的分子机器。现在，快速增长的蛋白质数据库拥有超过150,000个蛋白质结构。但是，要充分理解蛋白质作为机器的运行，必须了解其触发的功能动力学 - 蛋白质结构在被外部事件（例如配体对接，相邻蛋白质的运动）或电压或光强度变化的外部事件触发后及时演变。照片反应性蛋白质是一种特别令人兴奋的蛋白质类别，吸收光子后会产生机械能来调节生化过程和细胞行为。该项目将支持收敛研究，以测量两个照片反应蛋白中的光脉冲触发的时间分辨的会议变化：一种微生物视紫红质蛋白泵和称为光，氧和电压（LOV）敏感性域的光遗传学蛋白。 PI和Co-Pi在过去十年中的开发中实现了测量，该方法在非常高的磁场（8.6 Tesla）和频率（240 GHz）与位置定向诱变和使用基于GD（IIII III III III Centeragnentic Centers paramagnentic Centers paramagnentic Centements的频率（240 GHz）的频率（8.6 Tesla）和频率（240 GHz）结合在一起。该项目得到了生物科学局分子和细胞生物科学划分的分子生物物理学簇的支持。该奖项反映了NSF的法定任务，并被认为是通过基金会的智力优点和更广泛影响的评估来审查的审查标准，被认为是通过评估来支持的。

项目成果

Triggered Functional Dynamics of AsLOV2 by Time‐Resolved Electron Paramagnetic Resonance at High Magnetic Fields

高磁场下时间分辨电子顺磁共振触发 AsLOV2 的功能动力学

• DOI: 10.1002/ange.202212832
• 发表时间: 2023
• 期刊: Angewandte Chemie
• 作者: Maity, Shiny;Price, Brad D.;Wilson, C. Blake;Mukherjee, Arnab;Starck, Matthieu;Parker, David;Wilson, Maxwell Z.;Lovett, Janet E.;Han, Songi;Sherwin, Mark S.
• 通讯作者: Sherwin, Mark S.