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为生命提供动力的纳米级分子机器。由于结构生物学的一场革命，我们现在知道了超过15万种蛋白质的三维形状。然而，为了充分理解蛋白质作为机器的运作，人们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，相邻蛋白质的移动，或者电压或光强度的变化。一种特别令人兴奋的蛋白质，称为光响应蛋白质，可以在暴露于光时移动以控制细胞的行为。 这 研究将开发一种先进的磁共振形式--其中最常见的应用是磁共振成像（MRI）--来制作光响应蛋白质响应闪光的运动的“电影”。 “拍摄”蛋白质运动的能力将提供关键工具，以指导广泛的工程和优化蛋白质的重要应用，从可持续制造到用光控制神经。 该项目将通过主要研究者小组开发的“问题板”开展公共外联活动，吸引下一代科学家参与。蛋白质是我们所知的为生命提供动力的分子机器。快速增长的蛋白质数据库现在拥有超过15万个蛋白质结构。 然而，要完全理解蛋白质作为机器的运作，必须理解它们的触发功能动力学-蛋白质结构在被外部事件触发后如何及时演变，例如配体对接，相邻蛋白质的运动，或者电压或光强度的变化。 光响应蛋白是一类特别令人兴奋的蛋白质，在吸收光子后，产生机械能来调节生化过程和细胞行为。 这 项目 将支持收敛 研究 到 测量 的 在两种光响应蛋白中由光脉冲触发的时间分辨构象变化：微生物视紫红质质子泵和称为光、氧和电压（LOV）传感域的光遗传蛋白。 测量是通过一种独特的方法，在过去十年中由PI和co-PI开发，该方法将电子顺磁共振（EPR）在非常高的磁场（8.6特斯拉）和频率（240 GHz）与定点诱变和自旋标记结合起来，使用基于Gd（III）顺磁金属中心的化合物。该项目由生物科学理事会分子和细胞生物科学部的分子生物物理学小组支持。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.