Essential Role of Water and Lipids in G-Protein-Coupled Receptor Chemistry

水和脂质在 G 蛋白偶联受体化学中的重要作用

• 批准号: 1904125
• 负责人: Michael Brown
• 金额: $ 40万
• 依托单位: University of Arizona
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1904125&HistoricalAwards=false
• 关键词: Essential; Role; Water; Lipids; Protein

Rhodopsin is a visual protein found in the rods of the eye's retina, and is an archetype for the broad class of membrane proteins known as G-protein-coupled receptors (GPCRs). With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Michael Brown from the University of Arizona to explore how the interactions of this protein with lipids in membranes and with water control the visual signaling process. Although the atomic structures of proteins, including rhodopsin, are now known, the correspondence to their actions in lipid membranes remains an open question. This project combines experiment and theory to study how the flow of water into the protein affects the activation of rhodopsin within a cellular membrane environment. In addition, the project provides specialized training in the chemistry of membrane proteins and lipids to graduate students and postdoctoral fellows as well as undergraduate and high-school students. The new fundamental knowledge is disseminated not only through the published scientific literature, but also through workshops developed with the Arizona Flandreau Science Center. The project is further integrated with an outreach program involving Arizona's Science, Engineering, and Math Scholars program, focusing on promising students who are first in their family to attend college, from low-income households, and women and minorities. The work involves international collaborations and affords opportunities aimed at integrating and promoting human cooperation and global education. This research project characterizes important interactions involving membrane lipids and water that control the light activation of rhodopsin in the catalytic G-protein cycle of visual signaling. Because activation is a pH-dependent process, whether water goes into or out of the protein is crucial to the actual chemistry of the reaction mechanism. The project combines molecular spectroscopy with scattering methods through a synergetic interplay of experiment and theory. The research is intended to illuminate the hydration-mediated activation mechanism of rhodopsin, and explores how the membrane soft matter comprising lipids and water affects the process. Hydrophilic polymer osmolytes of low specificity are used to quantify the influx of water into the protein together with gravimetric methods, and reveal the influences of water on the activation of rhodopsin and its interactions with the cognate G-protein transducin. The project further addresses how the membrane lipids govern rhodopsin function in the cellular environment through the bilayer properties and interactions. The information from this study is intended to contribute new insights into the fundamental chemistry of the membrane components that are likely to govern G-protein-mediated biological signaling at the cellular level.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.

视紫红质是一种存在于视网膜视杆细胞中的视觉蛋白质，是一种被称为G蛋白偶联受体（GPCR）的膜蛋白的原型。 有了这个奖项，化学部的生命过程化学计划正在资助亚利桑那大学的Michael Brown博士，以探索这种蛋白质与膜中脂质和水的相互作用如何控制视觉信号传导过程。 尽管包括视紫红质在内的蛋白质的原子结构现在已经知道，但它们在脂膜中的作用的对应关系仍然是一个悬而未决的问题。该项目结合实验和理论来研究水流入蛋白质如何影响细胞膜环境中视紫红质的活化。 此外，该项目还为研究生和博士后研究员以及本科生和高中生提供膜蛋白和脂质化学方面的专门培训。新的基础知识不仅通过出版的科学文献传播，而且还通过与亚利桑那州弗兰德罗科学中心开发的研讨会传播。 该项目进一步与涉及亚利桑那州科学，工程和数学学者计划的外展计划相结合，重点关注家庭中第一个上大学的有前途的学生，来自低收入家庭，以及妇女和少数民族。这项工作涉及国际合作，并提供旨在整合和促进人类合作和全球教育的机会。该研究项目的特点涉及膜脂和水的重要相互作用，控制视紫红质的光激活在视觉信号的催化G蛋白循环。 由于活化是一个pH依赖性过程，水是否进入或离开蛋白质对反应机制的实际化学性质至关重要。 该项目通过实验和理论的协同作用将分子光谱学与散射方法相结合。 本研究旨在阐明水化介导的视紫红质激活机制，并探讨由脂质和水组成的膜软物质如何影响这一过程。 低特异性的亲水性聚合物渗透剂用于定量水流入蛋白质与重量法一起，并揭示水对视紫红质的活化及其与同源G-蛋白转导素的相互作用的影响。 该项目进一步解决了膜脂如何通过双层特性和相互作用来控制细胞环境中的视紫红质功能。 这项研究的信息旨在为可能在细胞水平上控制G蛋白介导的生物信号的膜成分的基本化学提供新的见解。该奖项反映了NSF的法定使命，并被认为值得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来支持。

项目成果

Nicotinamide phosphoribosyltransferase purification using SUMO expression system.

• DOI: 10.1016/j.ab.2020.113597
• 发表时间: 2020-06-01
• 期刊: Analytical biochemistry
• 影响因子: 2.9
• 作者: Molugu TR;Oita RC;Chawla U;Camp SM;Brown MF;Garcia JGN
• 通讯作者: Garcia JGN

Quantum Mechanical and Molecular Mechanics Modeling of Membrane-Embedded Rhodopsins

膜嵌入视紫红质的量子力学和分子力学建模

• DOI: 10.1007/s00232-019-00095-0
• 发表时间: 2019
• 期刊: The Journal of Membrane Biology
• 作者: Ryazantsev, Mikhail N.;Nikolaev, Dmitrii M.;Struts, Andrey V.;Brown, Michael F.
• 通讯作者: Brown, Michael F.

Phospholipid headgroups govern area per lipid and emergent elastic properties of bilayers.

磷脂头基控制着每个脂质的面积和双层的弹性特性。

• DOI: 10.1016/j.bpj.2022.09.005
• 发表时间: 2022
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Molugu,TrivikramR;Thurmond,RobinL;Alam,ToddM;Trouard,TheodoreP;Brown,MichaelF
• 通讯作者: Brown,MichaelF

Native mass spectrometry reveals the simultaneous binding of lipids and zinc to rhodopsin

• DOI: 10.1016/j.ijms.2020.116477
• 发表时间: 2021-02-01
• 期刊: INTERNATIONAL JOURNAL OF MASS SPECTROMETRY
• 影响因子: 1.8
• 作者: Norris, Carolanne E.;Keener, James E.;Marty, Michael T.
• 通讯作者: Marty, Michael T.

Biophysics of Membrane Stiffening by Cholesterol and Phosphatidylinositol 4,5-bisphosphate (PIP2)

• DOI: 10.1007/978-3-031-21547-6_2
• 发表时间: 2023-01-01
• 期刊: CHOLESTEROL AND PI(4,5)P2 IN VITAL BIOLOGICAL FUNCTIONS
• 作者: Doole, Fathima T.;Gupta, Sudipta;Brown, Michael F.
• 通讯作者: Brown, Michael F.