ENGINEERING RED-LIGHT ACTIVATED NUCLEOTIDE CYCLASES

工程红光激活核苷酸环化酶

基本信息

批准号：
8167818
负责人：
Mark Gomelsky
金额：
$ 3.1万
依托单位：
UNIVERSITY OF WYOMING
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-05-01 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8167818
关键词：
Adenylate Cyclase Affect Animal Disease Models Animal Model Binding Biomedical Research Cells Computer Retrieval of Information on Scientific Projects Database Cyclic AMP Diabetes Mellitus Engineering Funding Goals Grant Individual Institution Lasers Light Mammalian Cell Neuronal Plasticity Neurons Nucleotides Obesity Output Penetration Photons Photoreceptors Phototherapy Pilot Projects Protein Engineering Proteins Research Research Personnel Resolution Resources Role Source Tertiary Protein Structure Tissues United States National Institutes of Health Visible Radiation Work blood glucose regulation chromophore interest lipid metabolism spatiotemporal tissue/cell culture tool

Adenylate Cyclase Affect Animal Disease Models Animal Model Binding Biomedical Research Cells Computer Retrieval of Information on Scientific Projects Database Cyclic AMP Diabetes Mellitus Engineering Funding Goals Grant Individual Institution Lasers Light Mammalian Cell Neuronal Plasticity Neurons Nucleotides Obesity Output Penetration Photons Photoreceptors Phototherapy Pilot Projects Protein Engineering Proteins Research Research Personnel Resolution Resources Role Source Tertiary Protein Structure Tissues United States National Institutes of Health Visible Radiation Work blood glucose regulation chromophore interest lipid metabolism spatiotemporal tissue/cell culture tool

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项目摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Engineered photoregulated proteins have the potential to revolutionize biomedical research. In a photoregulated protein, a photon absorbed by a chromophore bound to a photoreceptor protein domain affects activity of an output domain. Visible light is practically harmless to mammalian cells, therefore, it can work as a highly specific, and affordable way to regulate protein activities. The spatiotemporal resolution that can be achieved by using photoregulated proteins is unprecedented as a laser beam can be focused not only on an individual cell but on a particular region of the cell. Engineered photoregulated proteins can be broadly used for activation (or inactivation) of proteins of interest in cell cultures, tissues and animal models. Thus far only blue-light photoreceptors have been used for protein engineering. Because of the short wavelengths of light they have low tissue penetration, which drastically limits their utility in animal models of disease. In contrast, bacteriophytochromes absorb red/far-red light, which has much higher tissue penetration capacity than blue light and is currently used in deep-tissue phototherapies. The objective of this application is to provide the proof of principle that a chromophore-binding module of bacteriophytochromes can be used for engineering of red/ far-red light regulated proteins. The goal of this pilot project is to engineer a red-light activated adenylate cyclase (cAMP synthase). The critical role of cAMP in controlling glucose and lipid metabolism as well as neuronal activity makes photoactivated adenylate cyclase a highly desired tool to study neuronal plasticity, progression of diabetes and obesity.

该副本是利用众多研究子项目之一由NIH/NCRR资助的中心赠款提供的资源。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构是对于中心，这不一定是调查员的机构。工程化的光蛋白蛋白具有彻底改变生物医学研究的潜力。在光子调节的蛋白质中，由发色团吸收到光感受器蛋白结构域的光子会影响输出结构域的活性。可见光实际上对哺乳动物细胞无害，因此，它可以作为一种调节蛋白质活性的高度特异性且负担得起的方式。可以通过使用光心调节蛋白来实现的时空分辨率是前所未有的，因为激光束不仅可以集中在单个细胞上，而且可以集中在单个细胞的特定区域上。工程化的光蛋白可以广泛用于细胞培养物，组织和动物模型中感兴趣的蛋白质的激活（或灭活）。迄今为止，仅将蓝光光感受器用于蛋白质工程。由于光的短波长，它们的组织渗透较低，这极大地限制了它们在疾病动物模型中的效用。相比之下，噬菌体色素吸收红色/远红色光，其组织渗透能力比蓝光高得多，目前用于深组织光疗法。该应用的目的是提供原理证明，表明拟菌植物色素的发色团结合模块可用于工程红色/远红色光调节蛋白。该试点项目的目的是设计红灯活化的腺苷环化酶（CAMP合酶）。 CAMP在控制葡萄糖和脂质代谢以及神经元活性中的关键作用使光活化的腺苷酸环化酶成为研究神经元可塑性，糖尿病进展和肥胖的高度期望的工具。