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.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 工程化的光调节蛋白质有可能彻底改变生物医学研究。在光调节蛋白质中，与光感受器蛋白结构域结合的发色团吸收的光子会影响输出结构域的活性。可见光对哺乳动物细胞几乎是无害的，因此，它可以作为一种高度特异性和负担得起的方式来调节蛋白质活性。通过使用光调节蛋白质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于单个细胞，而且可以聚焦于细胞的特定区域。工程化的光调节蛋白可广泛用于细胞培养物、组织和动物模型中感兴趣的蛋白的活化（或失活）。到目前为止，只有蓝光光受体被用于蛋白质工程。由于光的波长短，它们的组织穿透性低，这大大限制了它们在动物疾病模型中的应用。相比之下，细菌光敏色素吸收红光/远红光，其具有比蓝光高得多的组织穿透能力，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光/远红光调节蛋白的工程化的原理证明。该试验项目的目标是设计一种红光激活的腺苷酸环化酶（cAMP合成酶）。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得光活化腺苷酸环化酶成为研究神经元可塑性、糖尿病和肥胖症进展的高度期望的工具。