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: 8067163
• 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/8067163
• 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）： 近年来生物学领域最令人兴奋的技术发展之一是用光控制电活动的光化学方法的出现。我们在这个项目中的目标是通过生成赋予广泛的细胞表面受体和离子通道光敏感性的工具来开发一种广泛适用的方法来控制细胞功能的其他方面。为了应对这一挑战，我们将采用模块化方法，利用单个“通用光电开关”作为关键的光传感组件。该光电开关的核心包含小的可异构化偶氮苯部分，它分别响应 380 和 500 nm 的光而缩短和延长。我们将使用这种光开关来间接调节受体和通道活性，通过“适配器肽”，它包含一个“捕获域”，它识别光开关的短配置，但不识别长配置，以及一个配体域，其中包含靶细胞表面受体或离子通道的肽激活剂或抑制剂。所有衔接肽中的捕获结构域保持恒定，允许通过单个通用光开关进行控制，但配体结构域的性质是为了调节特定受体而定制的。将使用几种策略将接头肽的光依赖性捕获转化为受体激活或抑制。这些包括用二聚体光开关将衔接肽二聚化以激活生长因子受体，以及通过抗体系连的光开关将包括配体在内的衔接肽递送至G蛋白偶联受体。可以开发其他策略来激活不同类型的受体和离子通道。我们将测试通用光电开关方法对两个示例受体的有效性：脑源性生长因子 (BDNF) 的 TrkB 受体（受体酪氨酸激酶神经营养蛋白家族的成员）和神经肽 Y 的受体（GPCR） -型受体。产生一种对这些受体进行光敏调节的方法将能够以前所未有的精度检查它们在完整组织中的发育和神经功能中的作用，但更重要的是，它将证明一种强大的受体调节新技术的出现，可以应用存在已知肽配体的任何细胞表面蛋白。