A Model System for Sensor Proteins

传感器蛋白模型系统

基本信息

批准号：
6778334
负责人：
MICHAEL Anthony CUSANOVICH
金额：
$ 27.62万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-08-01 至 2006-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6778334
关键词：
Rhodospirillum X ray crystallography bacteriorhodopsins biological signal transduction chromophore computer simulation gene mutation nuclear magnetic resonance spectroscopy protein structure function rhodopsin site directed mutagenesis structural biology ultraviolet spectrometry western blottings

项目摘要

DESCRIPTION (provided by applicant): Photoactive yellow protein (PYP) is the prototype for a large family of proteins containing a structural motif referred to as the PAS domain. PAS domains function in a variety of signaling and regulatory roles in eucaryotes as well as procaryotes. Photoactive yellow protein (PYP) is a small soluble protein which appears to have a variety of signaling roles in purple bacteria. The goals of the proposed studies are four fold. 1. To elucidate the chemical and structural characteristics of the photocycle intermediates. 2.To understand the role of specific amino acid side chains in the kinetic and spectral properties of the ground state and intermediates. 3. To establish the mechanism(s) by which PYP signals to its reaction partners and utilizes the photocycle to function in vivo. 4. To establish the differences, in both functional and physico-chemical properties, among the three types of PYP identified to date. Structural, thermodynamic, kinetic and optical properties of the wild-type, site-specific mutants and variant chromophores will be studied. A variety of techniques will be utilized, including x-ray crystallography, NMR, UV/VIS spectroscopy, kinetic measurements in time domains ranging from femtoseconds to seconds, picosecond Coherent Anti-Stokes Raman Spectroscopy (CARS), picosecond time resolved CARS, and computer modeling.The significance of the proposed work is three fold. First, PYP will provide key insights which will help elucidate the role of PAS domains in signaling and regulation. This information will contribute directly to a better understanding of all of the members of the PAS domain family. Second, PYP offers the opportunity to understand the key elements in photoactive proteins - the spectral red-shift of the ground state protein and of key intermediates, how photoisomerization can be used to drive function, and the structure of photocycle intermediates. This level of understanding will complement work with membrane-bound photoactive proteins (bacteriorhodopsin, sensory rhodopsin and rhodopsin) and will provide information not available with these systems. Third, it is possible that an understanding of PYP and photoactive proteins in general will contribute to the development of photobioelectronic switching devices which will have applications in the next generation of small, fast, low power electronics

描述（由申请人提供）：光活性黄蛋白（PYP）是大型蛋白质的原型，该蛋白质包含一个称为PAS结构域的结构基序。 PAS结构域在各种信号传导和调节作用中发挥作用，在桉树和procaryotes中起作用。光活性黄蛋白（PYP）是一种小型可溶性蛋白，在紫色细菌中似乎具有多种信号传导作用。拟议的研究的目标是四倍。 1。阐明光循环中间体的化学和结构特征。 2.了解特定氨基酸侧链在基态和中间体的动力学和光谱特性中的作用。 3。建立PYP向其反应伙伴信号并利用光圈在体内发挥作用的机制。 4。在迄今为止确定的三种类型的PYP中，在功能和物理化学特性中建立差异。将研究野生型，位点特异性突变体和变体生色团的结构，热力学，动力学和光学性质。 A variety of techniques will be utilized, including x-ray crystallography, NMR, UV/VIS spectroscopy, kinetic measurements in time domains ranging from femtoseconds to seconds, picosecond Coherent Anti-Stokes Raman Spectroscopy (CARS), picosecond time resolved CARS, and computer modeling.The significance of the proposed work is three fold.首先，PYP将提供关键的见解，这将有助于阐明PAS域在信号传导和调节中的作用。这些信息将直接有助于更好地理解PAS域家族的所有成员。其次，PYP提供了了解光活动蛋白的关键要素 - 基态蛋白质和关键中间体的光谱，如何使用光异构化来驱动功能以及光循环中间体的结构。这种理解水平将与结合膜结合的光活性蛋白（细菌紫红素，感觉视紫红质和视紫红质）的工作相辅相成，并将提供这些系统无法获得的信息。第三，对PYP和光活性蛋白的理解可能会有助于照射电源开关设备的开发，该设备将在下一代小型，快速，低功率电子产品中应用

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Time-resolved single tryptophan fluorescence in photoactive yellow protein monitors changes in the chromophore structure during the photocycle via energy transfer.

光敏黄色蛋白中的时间分辨单色氨酸荧光通过能量转移监测光循环期间发色团结构的变化。

DOI：
10.1021/bi051448l
发表时间：
2005
期刊：
Biochemistry.
影响因子：
0
作者：
Otto,Harald;Hoersch,Daniel;Meyer,TerryE;Cusanovich,MichaelA;Heyn,MaartenP
通讯作者：
Heyn,MaartenP

pH Dependence of the photocycle kinetics of the E46Q mutant of photoactive yellow protein: protonation equilibrium between I1 and I2 intermediates, chromophore deprotonation by hydroxyl uptake, and protonation relaxation of the dark state.

光活性黄色蛋白 E46Q 突变体的光循环动力学的 pH 依赖性：I1 和 I2 中间体之间的质子化平衡、羟基摄取引起的发色团去质子化以及暗态的质子化弛豫。

DOI：
10.1021/bi034315d
发表时间：
2003
期刊：
Biochemistry.
影响因子：
0
作者：
Borucki,Berthold;Otto,Harald;Joshi,ChandraP;Gasperi,Chiara;Cusanovich,MichaelA;Devanathan,Savitha;Tollin,Gordon;Heyn,MaartenP
通讯作者：
Heyn,MaartenP

Thermochromatium tepidum photoactive yellow protein/bacteriophytochrome/diguanylate cyclase: characterization of the PYP domain.

Thermochromatium tepidum 光敏黄色蛋白/细菌光敏色素/二鸟苷酸环化酶：PYP 结构域的表征。