ENGINEERING RED-LIGHT ACTIVATED NUCLEOTIDE CYCLASES

工程红光激活核苷酸环化酶

• 批准号: 8359737
• 负责人: Mark Gomelsky
• 金额: $ 3.42万
• 依托单位: UNIVERSITY OF WYOMING
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2012-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8359737
• 关键词: Adenylate Cyclase; Affect; Animal Model; Binding; Bioinformatics; Biomedical Research; Cells; Cyclic AMP; Engineering; Enzymes; Funding; Goals; Grant; Individual; Lasers; Light; Mammalian Cell; Modeling; National Center for Research Resources; Neurons; Nucleotides; Output; Penetration; Photons; Photoreceptors; Phototherapy; Pilot Projects; Principal Investigator; Protein Dynamics; Protein Engineering; Proteins; Research; Research Infrastructure; Resolution; Resources; Role; Source; Tertiary Protein Structure; Tissues; United States National Institutes of Health; Work; Wyoming; chromophore; cost; design; glucose metabolism; interest; lipid metabolism; protein structure; spatiotemporal; tissue/cell culture

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. 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. Infrared light is harmless to mammalian cells, therefore, it can work as a highly specific, and cheap 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. Bacteriophytochromes absorb red/near infrared 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/near infrared light regulated proteins. The goal of this pilot project is to engineer a near infrared light activated adenylate cyclase (cAMP synthase). The critical role of cAMP in controlling glucose and lipid metabolism as well as neuronal activity makes adenylate cyclase a highly desired target. The design of photoregulated enzymes relies heavily on computationally-intensive bioinformatics approaches that involve analysis and modeling of protein structures and dynamics.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 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。红外光对哺乳动物细胞无害，因此，它可以作为一种高度特异性和廉价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。细菌光敏色素吸收红光/近红外光，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/近红外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近红外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腺苷酸环化酶成为高度期望的靶标。光调控酶的设计在很大程度上依赖于计算密集型生物信息学方法，包括蛋白质结构和动力学的分析和建模。