ENGINEERING RED-LIGHT ACTIVATED NUCLEOTIDE CYCLASES

工程红光激活核苷酸环化酶

• 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
• 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

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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 工程光调节蛋白有可能彻底改变生物医学研究。在光调节蛋白中，被与光感受器蛋白结构域结合的发色团吸收的光子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，它可以作为一种高度特异性且经济实惠的调节蛋白质活性的方法。使用光调节蛋白可以实现的时空分辨率是前所未有的，因为激光束不仅可以聚焦在单个细胞上，而且可以聚焦在细胞的特定区域上。工程光调节蛋白可广泛用于激活（或失活）细胞培养物、组织和动物模型中感兴趣的蛋白质。迄今为止，只有蓝光光感受器被用于蛋白质工程。由于光波长短，它们的组织穿透性低，这极大地限制了它们在疾病动物模型中的应用。相比之下，细菌光敏色素吸收红/远红光，其组织穿透能力比蓝光高得多，目前用于深层组织光疗。本申请的目的是提供细菌光敏色素的发色团结合模块可用于红/远红光调节蛋白工程的原理证明。该试点项目的目标是设计一种红光激活的腺苷酸环化酶（cAMP 合酶）。 cAMP 在控制葡萄糖和脂质代谢以及神经元活动中的关键作用使得光激活腺苷酸环化酶成为研究神经元可塑性、糖尿病和肥胖进展的非常理想的工具。