Genome-wide Investigation of PDZ Domain Specificity

PDZ 结构域特异性的全基因组研究

• 批准号: 7935593
• 负责人: GAVIN MACBEATH
• 金额: $ 11.12万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7935593
• 关键词: Address; Biochemical; Biological; Biological Assay; Catalytic Domain; Cell Nucleus; Cell Polarity; Cell membrane; Cells; Collection; Data; Databases; Development; Disease; Endothelial Cells; Epithelial; Evaluation; Event; Fluorescence Polarization; Foundations; Future; Gel Chromatography; Hand; Human; Individual; Internet; Investigation; Knockout Mice; Ligands; Location; Mediating; Methods; Microarray Analysis; Molecular; Mus; Neurons; Neuropathy; PDZ protein; Peptides; Peripheral; Phase; Phenotype; Physiological; Play; Process; Property; Protein Microchips; Proteins; Publishing; Randomized; Receptor Protein-Tyrosine Kinases; Relative (related person); Research Personnel; Role; Sensorineural Hearing Loss; Sensory; Signal Transduction; Signaling Protein; Solutions; Specificity; Substrate Interaction; Synapses; Usher Syndrome; computerized data processing; enzyme substrate; genome-wide; mouse genome; myelinopathy; numb protein; postsynaptic; presynaptic; programs; protein complex; protein transport; research study

DESCRIPTION (provided by applicant): Most eukaryotic proteins that receive and process signals are constructed from a combination of interaction and catalytic domains. Among the many interaction domains identified in the past decade, PDZ's are one of the most frequently encountered. They are often found in combination with other interaction modules and play a role in directing the specificity of receptor tyrosine kinase-mediated signaling, in establishing cell polarity, in directing protein trafficking, and in coordinating synaptic signaling. Their importance is underscored by the severe neuronal and developmental phenotypes observed in PDZ knockout mice and by their implication in human congenital diseases like Usher syndrome and Dejerine-Sottas neuropathy. The enormous diversity of PDZ function is manifest in their abundance; there are over 250 PDZ's encoded in the mouse genome. To understand their individual roles, it is necessary first to define their recognition properties in a comprehensive and relevant fashion. Previous efforts to define PDZ selectivity have focused on only a few domains and have relied on collections of randomized peptides, rather than on physiological ligands. In order to provide a genome-wide understanding of PDZ function, the selectivity of every PDZ encoded in the mouse genome will be investigated relative to a large collection of genomically-encoded ligands using protein microarray technology. The aims of this proposal are: (1) to clone, express, and purify every mouse PDZ domain; (2) to screen every PDZ with every PDZ to reveal putative PDZ-PDZ interactions; (3) to screen every PDZ with 222 genomically-encoded peptide ligands to identify putative PDZ-protein interactions; (4) to investigate the physiological relevance of a subset of predicted interactions biochemically; and (5) to construct a relational database that integrates protein microarray data with published information on PDZ domains. Collectively, these efforts should provide the necessary foundation to understand PDZ function on a genome-wide level and aid future efforts to intervene appropriately when PDZ function goes awry.

描述（由申请人提供）： 大多数接收和处理信号的真核蛋白质是由相互作用和催化结构域的组合构建的。在过去十年中发现的许多相互作用域中，PDZ 是最常遇到的域之一。它们通常与其他相互作用模块结合使用，并在指导受体酪氨酸激酶介导的信号传导的特异性、建立细胞极性、指导蛋白质运输和协调突触信号传导方面发挥作用。 在 PDZ 敲除小鼠中观察到的严重神经元和发育表型及其对人类先天性疾病（如 Usher 综合征和 Dejerine-Sottas 神经病）的影响凸显了它们的重要性。 PDZ 功能的巨大多样性体现在其丰富性上；小鼠基因组中编码了超过 250 个 PDZ。要了解他们的个人角色，有必要首先以全面且相关的方式定义他们的识别属性。先前定义 PDZ 选择性的努力仅集中于少数几个域，并且依赖于随机肽的集合，而不是生理配体。 为了提供对 PDZ 功能的全基因组理解，将使用蛋白质微阵列技术研究小鼠基因组中编码的每个 PDZ 相对于大量基因组编码配体的选择性。该提案的目的是：（1）克隆、表达和纯化每个小鼠PDZ结构域； (2) 筛选每个PDZ与每个PDZ以揭示假定的PDZ-PDZ相互作用； (3) 用 222 个基因组编码的肽配体筛选每个 PDZ，以鉴定假定的 PDZ-蛋白质相互作用； (4) 通过生化研究预测的相互作用子集的生理相关性； (5) 构建一个关系数据库，将蛋白质微阵列数据与已发表的 PDZ 结构域信息相结合。总的来说，这些努力应该为在全基因组水平上理解 PDZ 功能提供必要的基础，并有助于未来在 PDZ 功能出现问题时进行适当干预。