A universal photoswitch system for optical control of neuronal receptors

用于神经元受体光学控制的通用光电开关系统

• 批准号: 8255457
• 负责人: RICHARD H KRAMER
• 金额: $ 30.39万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-22 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8255457
• 关键词: Antibodies; Biochemical; Biology; Brain-Derived Neurotrophic Factor; Cell Surface Proteins; Cell Surface Receptors; Cell physiology; Development; Effectiveness; Family; G Protein-Coupled Receptor Genes; G-Protein-Coupled Receptors; Goals; Growth Factor Receptors; Health; Ion Channel; Ligands; Light; Methods; Nature; Neurons; Neurophysiology - biologic function; Neurotrophic Tyrosine Kinase Receptor Type 2; Optics; Peptides; Photophobia; Receptor Activation; Receptor Inhibition; Receptor Protein-Tyrosine Kinases; Regulation; Role; System; Techniques; Testing; Tissues; Translating; azobenzene; brain-derived growth factor; combinatorial; inhibitor/antagonist; meetings; member; neuropeptide Y; neurotrophic factor; receptor; response; tool

DESCRIPTION (provided by applicant): One of the most exciting technical developments in biology in recent years is the emergence of photochemical methods for controlling electrical activity with light. Our goal in this project is to develop a broadly applicable method for controlling other aspects of cellular function by generating tools for conferring light sensitivity on a broad range of cell surface receptors and ion channels. To meet this challenge we will use a modular approach, utilizing a single "Universal Photoswitch" as the key light-sensing component. The photoswitch contains at its core the small isomerizable azobenzene moiety, which shortens and lengthens in response to 380 and 500 nm light, respectively. We will use this photoswitch to indirectly regulate receptor and channel activity, through an "adapter peptide", which contains a "capture domain", which recognizes the short, but not the long configuration of the photoswitch, and a ligand domain, which contains a peptide activator or inhibitor of the targeted cell surface receptor or ion channel. The capture domain is kept constant among all adapter peptides, allowing control by a single Universal Photoswitch, but the nature of ligand domain is tailored to regulate a specific receptor. Several strategies will be used to translate light-dependent capture of the adapter peptide into receptor activation or inhibition. These include dimerizing the adapter peptide with a dimeric photoswitch to activate growth factor receptors, and delivering the adapter peptide, including the ligand, to a G-protein coupled receptor via an antibody-tethered photoswitch. Other strategies may be developed to activate different types of receptors and ion channels. We will test the effectiveness of the Universal Photoswitch approach on two example receptors: the TrkB receptor for brain-derived growth factor (BDNF), a member of the neurotrophin family of receptor tyrosine kinases, and the receptor for neuropeptide Y, which is a GPCR-type receptor. Generating a method for light-sensitive regulation of these receptors will allow examination of their roles in development and neural function in intact tissue with unprecedented precision, but more importantly, it will demonstrate the emergence of a powerful new technique for receptor regulation that can be applied to any cell surface protein for which a known peptide ligand exists.

描述（由申请人提供）： 近年来，生物学上最令人兴奋的技术发展之一是光化学方法的出现，用于控制光线活性。我们在该项目中的目标是通过生成在广泛的细胞表面受体和离子通道上赋予光灵敏度的工具来开发一种可控制细胞功能其他方面的广泛适用方法。为了应对这一挑战，我们将使用模块化方法，利用单个“通用Photoswitch”作为关键的光感应组件。 Photoswitch在其核心中包含了小的异构偶氮苯一部分，该部分分别缩短并延长了380和500 nm光的响应。我们将使用此照片开关通过“衔接肽”间接调节受体和通道活动，该肽包含一个“捕获结构域”，该肽识别photoswitch的短构型，但不识别Photoswitch的长配置，以及一个包含靶标细胞表面受体或ion ion ion Channel的肽激活剂或抑制剂的配体域。在所有衔接肽中，捕获结构域保持恒定，从而可以通过单个通用照片开关进行控制，但是配体域的性质是量身定制的，以调节特定的受体。将使用几种策略将衔接肽的光依赖捕获转化为受体激活或抑制作用。这些包括用二聚体的照片开关将衔接肽二聚二聚体以激活生长因子受体，并通过抗体螺旋螺旋射线开关将衔接肽（包括配体）传递到G蛋白偶联受体中。可以开发其他策略来激活不同类型的受体和离子通道。我们将在两个示例受体上测试通用照片开关方法的有效性：脑衍生生长因子的TRKB受体（BDNF），受体酪氨酸激酶的神经营养蛋白家族的成员和神经肽Y的受体，是GPCR型受体。生成一种对这些受体的光敏调节的方法，将允许以前所未有的精度来检查其在完整组织中的发育和神经功能中的作用，但更重要的是，它将证明可以将有力的受体调节的新技术出现在任何可应用于任何已知肽座的细胞表面蛋白质上。