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着很大的希望，但它们的应用受到蛋白质工程策略的限制，这些策略是劳动密集型的，需要一定水平的生物化学和细胞工程复杂性，而这在许多生物学实验室中是不可用的。事实上，尽管光遗传学的潜力已经得到了很大的发展，但令人惊讶的是，很少有基因编码的光响应蛋白被描述。有没有可能设计出一种光遗传学蛋白质工程策略，这种策略非常简单，生物学家可以充当自己的蛋白质工程师？在这方面，我们已经制定了一个潜在的一般战略，从100岁的Michaelis Menten方程中汲取灵感。这种方法提供了一个光激活的cofilin（光介导的细胞运动）和一个光激活的bax（光介导的细胞死亡）。我们将准备另外三种光响应蛋白，以探索这种策略的范围和局限性。要获得的三个构建体，连同迄今为止开发的两个构建体，是已知调节线粒体行为的蛋白质大家族的代表。几种神经系统疾病（帕金森氏病、亨廷顿病、阿尔茨海默氏病和腓骨肌萎缩症2A型）显示线粒体动力学缺陷，包括融合、分裂、运输和周转。最近的研究表明，通过改变线粒体动力学可以改善特定的疾病表型。我们将通过研究光响应蛋白以光依赖方式调节线粒体行为的能力来探索这一前提。