Optogenetic Mitochondria-Directed Proteins

 描述(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，尽管光遗传工具在生物学和医学上有希望，但它们的现成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，需要达到许多生物实验室所不具备的生化和细胞工程的复杂程度。事实上，尽管光遗传学的潜力已经有了很多，但令人惊讶的是，很少有基因编码的光响应蛋白被描述出来。有没有可能设计出一种如此简单的光遗传蛋白质工程策略，以至于生物学家可以充当自己的蛋白质工程师？在这方面，我们开发了一种潜在的一般策略，该策略的灵感来自于已有100年历史的米凯利斯·曼腾方程。这种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)。我们将另外准备三个光响应蛋白，以探索这一策略的范围和局限性。将获得的三个结构，与迄今开发的两个结构一起，是已知的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(帕金森氏症、亨廷顿氏症、阿尔茨海默氏症和夏科-玛丽-牙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，有可能通过改变线粒体动力学来改善特定的疾病表型。我们将通过检测被研究的光响应蛋白以光依赖的方式调节线粒体行为的能力来探索这一前提。

 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.