Optogenetic Mitochondria-Directed Proteins

光遗传学线粒体定向蛋白

• 批准号: 8967476
• 负责人: DAVID S. LAWRENCE
• 金额: $ 22.8万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8967476
• 关键词: Address; Alzheimer' s Disease; Animal Model; Attention; Behavior; Biochemical; Biochemistry; Biological; Biology; Breathing; Cell Death; Cells; Centenarian; Charcot-Marie-Tooth Disease; Databases; Defect; Dependence; Development; Disease; Drug Delivery Systems; Engineering; Enzymatic Biochemistry; Equation; Event; Funding; Grant; Huntington Disease; Light; Malignant Neoplasms; Mediating; Medicine; Methods; Mitochondria; Organelles; Organism; Parkinson Disease; Protein Engineering; Protein Family; Proteins; PubMed; Research Personnel; Role; Science; Series; Staging; United States National Institutes of Health; Vitamin B 12; analog; cell behavior; cell motility; cellular engineering; cofilin; design; disease phenotype; inhibitor/antagonist; nervous system disorder; optogenetics; public health relevance; small molecule; tool

 DESCRIPTION (provided by applicant): The ability to control the biochemistry of cells and organisms with light has elicited widespread attention. However, in spite of the promise that optogenetic tools hold for biology and medicine, their ready application is constrained by protein engineering strategies that are labor intensive and require a level of biochemical and cellular engineering sophistication that is not available in many biology labs. Indeed, although much has been made of the potential of optogenetics, surprisingly few genetically encoded light-responsive proteins have been described. Is it possible to devise an optogenetic protein engineering strategy that is so straightforward that biologists can serve as their own protein engineers? In this regard, we have developed a potentially general strategy that draws its inspiration from the 100-year-old Michaelis Menten equation. This approach has furnished a light-activatable cofilin (light-mediated cell motility) and a light-activatable bax (light-mediate cell death). We will prepare three additional light-responsive proteins in order to explore the scope and limitations of this strategy. The three constructs to be acquired, in conjunction with the two developed to date, are representatives of a large family of proteins known to modulate mitochondrial behavior. Several neurological diseases (Parkinson's, Huntington's, Alzheimer's, and Charcot-Marie-Tooth type 2A) display defects in mitochondrial dynamics, including fusion, fission, transport, and turnover. Recent studies have suggested that it may be possible to ameliorate specific disease phenotypes by altering mitochondrial dynamics. We will explore this premise by examining the ability of the light-responsive proteins under study to modulate mitochondrial behavior in a light-dependent fashion.

 描述（由申请人提供）：用光控制细胞和生物体的生物化学的能力已经引起了广泛的关注。然而，尽管光遗传学工具对生物学和医学有希望，但它们的现成应用受到蛋白质工程策略的限制。光遗传学是劳动密集型的，并且需要一定程度的生化和细胞工程复杂性，而这是许多生物实验室所不具备的。事实上，尽管光遗传学的潜力已经得到了很大的发挥，但令人惊讶的是，基因编码的技术却很少。是否有可能设计出一种如此简单的光遗传学蛋白质工程策略，使生物学家可以充当自己的蛋白质工程师？在这方面，我们从 100 个例子中汲取了灵感，开发了一种潜在的通用策略。该方法提供了光激活的 cofilin（光介导的细胞运动）和光激活的 bax（光介导的细胞死亡）。额外的光响应蛋白，以探索该策略的范围和局限性。要获得的三个构建体以及迄今为止开发的两个构建体是已知调节几种神经系统疾病的蛋白质大家族的代表。 （帕金森病、亨廷顿病、阿尔茨海默病和腓骨肌萎缩症 2A 型）在线粒体动力学方面表现出缺陷，包括融合、裂变、运输和周转。最近的研究表明，这可能是可能的。通过改变线粒体动力学来改善特定的疾病表型我们将通过检查所研究的光响应蛋白以光依赖性方式调节线粒体行为的能力来探索这一前提。