International Collaboration in Chemistry on Control of Excited State Proton Transfer in GFP

控制 GFP 中激发态质子转移的国际化学合作

基本信息

批准号：
EP/I003304/1
负责人：
Jasper Van Thor
金额：
$ 61.95万
依托单位：
Imperial College London
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2011
资助国家：
英国
起止时间：
2011 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FI003304%2F1
关键词：
International Collaboration Chemistry Control Excited

项目摘要

The 2008 Nobel Prize recognized the revolutionary biological applications of the green fluorescent protein (GFP) and related fluorescent proteins as tools for monitoring protein localization and trafficking in cells and tissues. However, the prototype GFP, from the jellyfish Aequorea victoria, is also unique in exhibiting highly efficient (>80%) proton transfer following absorption of a photon. Proton transfer contributes to most biological reactions, but typically takes place on time scales that are difficult to observe directly. The ability to trigger proton transfer with a femtosecond light pulse thus yields unequalled opportunities for understanding how proteins control this ubiquitous process. Our proposed investigations proposal is hypothesis-driven and aims to understand the dynamics and control of the ultrafast excited state proton transfer (ESPT) reaction. In particular, we hope to identify low frequency promoting vibrations that form an essential element of theoretical treatments of proton transfer. We present preliminary measurements of low frequency vibrational coherences that occur within the duration of the ESPT reaction. Control of ESPT in GFP is currently fervidly discussed and we propose to help resolve outstanding issues such as the enthalpic and entropic reaction barriers, the role of chromophore cooling, low frequency mode identification and assignment, and heterogeneity and interconversions of the protein-chromophore structure. Ultrafast kinetics, vibrational coherence spectroscopy (VCS), transient visible-visible and visible-infrared absorption spectroscopy, femtosecond stimulated Raman spectroscopy (FSRS), and coherent control spectroscopy will be used to dissect the microscopic details. Particular emphasis will be placed on temperature dependent studies. Further development of high level computational chemistry calculations that include the protein environment will aid in interpretation of the spectroscopic results. In turn, the experimental results will provide an essential calibration for the computational methods.

2008 年诺贝尔奖表彰了绿色荧光蛋白 (GFP) 和相关荧光蛋白作为监测细胞和组织中蛋白质定位和运输的工具的革命性生物学应用。然而，来自维多利亚多管发光水母的 GFP 原型在吸收光子后表现出高效 (>80%) 质子转移方面也是独一无二的。质子转移有助于大多数生物反应，但通常发生在难以直接观察的时间尺度上。因此，利用飞秒光脉冲触发质子转移的能力为了解蛋白质如何控制这一普遍过程提供了无与伦比的机会。我们提出的研究方案是假设驱动的，旨在了解超快激发态质子转移（ESPT）反应的动力学和控制。特别是，我们希望确定低频促进振动，这是质子转移理论治疗的基本要素。我们对 ESPT 反应期间发生的低频振动相干性进行了初步测量。目前，对 GFP 中 ESPT 的控制进行了热烈的讨论，我们建议帮助解决一些突出的问题，例如熵和熵反应障碍、发色团冷却的作用、低频模式识别和分配以及蛋白质-发色团结构的异质性和相互转换。超快动力学、振动相干光谱（VCS）、瞬态可见光和可见红外吸收光谱、飞秒受激拉曼光谱（FSRS）和相干控制光谱将用于剖析微观细节。将特别强调温度依赖性研究。包括蛋白质环境在内的高级计算化学计算的进一步发展将有助于解释光谱结果。反过来，实验结果将为计算方法提供必要的校准。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Ultrafast vibrational dynamics of parallel excited state proton transfer reactions in the Green Fluorescent Protein

绿色荧光蛋白中平行激发态质子转移反应的超快振动动力学

DOI：
10.1016/j.vibspec.2012.03.005
发表时间：
2012-09-01
期刊：
Vibrational Spectroscopy
影响因子：
2.5
作者：
J. J. Thor;C. Lincoln;B. Kellner;K. Bourdakos;Lee M. Thompson;M. Bearpark;P. Champion;J. Sage
通讯作者：
J. Sage

Room temperature crystal structure of the fast switching M159T mutant of the fluorescent protein dronpa.

荧光蛋白 dronpa 快速开关 M159T 突变体的室温晶体结构。