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.

蛋白质是为我们所知的生命提供动力的纳米分子机器，得益于结构生物学的革命，我们现在知道了超过 150,000 种蛋白质的三维形状。人们必须了解它们在被外部事件触发后如何移动，例如附着到特定分子、邻近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，称为光响应蛋白质，可以通过移动来控制。细胞在暴露于环境时的行为这项研究将开发一种先进的磁共振技术，其中最常见的应用是磁共振成像（MRI），以制作光响应蛋白响应闪光的运动“电影”。 “薄膜”蛋白质运动将提供关键工具来指导广泛的努力，以设计和优化蛋白质，以实现从可持续制造到用光控制神经的重要应用。该项目将通过开发的“Questboard”通过公共宣传活动吸引下一代科学家。 PI 的小组是分子机器。快速增长的蛋白质数据库现在拥有超过 150,000 个蛋白质结构。然而，要充分了解蛋白质作为机器的运作，我们必须了解它们触发的功能动力学——蛋白质结构如何及时演化。由配体对接、邻近蛋白质的运动或电压或光强度的变化等外部事件触发的光响应蛋白是一类特别令人兴奋的蛋白质，它们在吸收光子后产生机械能来调节生化过程。和细胞行为。将支持聚合研究，以测量两种光响应蛋白中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）传感域的光遗传学蛋白。测量已启用。 PI 和 co-PI 在过去十年中开发了一种独特的方法，该方法结合了极高磁场（8.6 特斯拉）和频率（240 GHz），使用基于 Gd (III) 顺磁金属中心的化合物进行定点诱变和自旋标记。该项目得到生物科学理事会分子和细胞生物科学部分子生物物理学集群的支持。该奖项反映了 NSF 的法定使命。通过使用基金会的智力优点和更广泛的影响审查标准进行评估，并被认为值得支持。

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: 2023-02
• 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.