Structural and Functional Studies of Teneurins: A bacterial toxin homolog in human

Teneurins 的结构和功能研究：人类细菌毒素同系物

• 批准号: 10237184
• 负责人: Demet Arac-Ozkan
• 金额: $ 32.4万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10237184
• 关键词: Adhesions; Affinity; Alternative Splicing; Amino Acids; Architecture; Bacterial Toxins; Binding; Biochemical; Biological; Biological Assay; Biophysics; Brain; C-terminal; Cell Adhesion; Cell Adhesion Molecules; Cell Aggregation; Cell physiology; Cell surface; Cells; Cellular biology; Communication; Complement; Complex; Coupling; Cryoelectron Microscopy; Cues; Cyclic AMP; Data; Dependence; Development; Dimerization; Disease; Electron Microscopy; Embryonic Development; Encapsulated; Excitatory Synapse; Exhibits; Family; G-Protein-Coupled Receptors; Goals; Homodimerization; Homologous Gene; Human; Immunoglobulin Domain; Inhibitory Synapse; Integral Membrane Protein; Knowledge; Laboratories; Lasers; Ligands; Link; Mechanics; Mediating; Membrane Proteins; Methods; Molecular; Monitor; Mutagenesis; Mutate; Negative Staining; Nervous system structure; Neurons; Organ; Organism; Physiological; Play; Protein Family; Protein Isoforms; Proteins; RNA Splicing; Reporting; Research; Resolution; Role; Rosaniline Dyes; Sequence Analysis; Signal Transduction; Site; Structure; Structure-Activity Relationship; Surface; Synapses; System; Testing; Tissues; Toxin; Variant; Visualization; alpha Toxin; alpha-latrotoxin receptor; analytical ultracentrifugation; anticancer research; axon guidance; base; cardiogenesis; crosslink; experimental study; extracellular; follow-up; human disease; in vivo; insight; intercellular communication; interdisciplinary approach; light scattering; malignant neurologic neoplasms; member; mutant; nervous system disorder; neural circuit; novel; novel strategies; organizational structure; protein function; relating to nervous system; success; synaptic function; synaptogenesis

Project Summary The interplay between cellular adhesion and cellular signaling is essential for the development of all organs such as the brain, and for the functioning of systems such as the nervous systems. Teneurins (TEN1-4) are a poorly understood family that mediates intercellular communication. They have essential roles in embryonic development and neural circuit-wiring; and are linked to numerous human diseases including neurological disorders and cancers. TENs are type-II membrane proteins with large C-terminal extracellular regions (ECR) that majorly exhibit no identifiable domains. The ECR mediates trans-cellular heterophilic interaction of TENs with Latrophilins(LPHN1-3), a family of G-Protein Coupled Receptors; to regulate synapse function. The ECR also mediates trans-cellular homophilic interaction of TENs with themselves to instruct neural circuit-wiring. However, the molecular mechanisms underlying TEN action remains poorly understood majorly due to the lack of structural information on the ECR. We recently laid the groundwork by determining the high-resolution cryo-EM structure of the TEN2 ECR and revealed a surprising homology to bacterial Tc-toxins. We also showed that an alternatively spliced insert acts as a switch to regulate LPHN binding and other TEN functions such as synapse formation. The ultimate goal of the research proposed in this application is to understand the mechanical details of various TEN functions that are mediated by its ECR. We propose three Specific Aims that are based on the major unknowns in TEN function and a Follow-up Aim to perform structure/function relationship studies: First, we aim to understand the molecular details of the TEN/LPHN interaction. Second, we aim to understand the molecular determinants for the trans-homodimerization of TEN. Third, we aim to reveal whether TEN functions via autoproteolysis similar to bacterial toxins. Then, we aim to use the information from the first three aims to study TEN function in synapse formation assays. This research has a multi-disciplinary approach where the structural and functional data performed in the PI's lab range from electron microscopy, biophysical and biochemical methods, neuronal assays to cell-biology and is complemented by the expertise provided or performed by the laboratories of close collaborators. The proposed experiments will build on exciting results, including the very unusual TEN2 structure, surprising involvement of alternative splicing in TEN function, key advances in the purification of all needed TEN fragments, and the observation of proteolytic products. We expect that this research will provide critical insights into the mechanistic details of TEN function, helping to establish novel principles on intercellular communication that are vital for numerous cellular functions.

项目摘要 细胞粘附和细胞信号之间的相互作用对于所有器官的发展至关重要 例如大脑，以及神经系统等系统的功能。 Teneurins（Ten1-4）是 介导细胞间交流的家庭了解不足。他们在胚胎中具有重要的作用 开发和神经电路系；并与包括神经系统在内的众多人类疾病有关 疾病和癌症。 TENS是II型膜蛋白，具有较大的C末端细胞外区域（ECR） 这主要显示出无法识别的域。 ECR介导Tens的跨细胞异质相互作用 与ltrophilins（LPHN1-3），G蛋白偶联受体家族；调节突触功能。 ECR 还介导了数十个与自身的跨细胞均电相互作用，以指导神经电路。 但是，十项作用的分子机制主要是由于 缺乏有关ECR的结构信息。我们最近通过确定高分辨率来奠定基础 TEN2 ECR的冷冻EM结构，并揭示了与细菌TC毒素具有令人惊讶的同源性。我们也是 表明替代剪接的插入物是调节LPHN结合和其他十个功能的开关 例如突触形成。本应用程序提出的研究的最终目标是了解 由其ECR介导的各种十种功能的机械细节。我们提出了三个具体目标 基于十个功能中的主要未知数和执行结构/功能的后续目标 关系研究：首先，我们旨在了解十/LPHN相互作用的分子细节。第二， 我们的目标是了解十个跨二胺化的分子决定因素。第三，我们的目标是 揭示通过类似细菌毒素类似的自动溶解的十个功能。然后，我们的目的是使用 前三个旨在研究突触形成测定中的十个功能的信息。这项研究有 多学科的方法，在PI实验室中执行的结构和功能数据范围从 电子显微镜，生物物理和生化方法，神经元测定到细胞生物学，IS 与密切合作者实验室提供或执行的专业知识相辅相成。提议 实验将基于令人兴奋的结果，包括非常不寻常的TEN2结构，令人惊讶的参与 十个功能中的替代剪接，纯化所有必需的十个片段的关键进展，以及 观察蛋白水解产物。我们希望这项研究将为您提供关键见解 十个功能的机械细节，有助于建立有关细胞间交流的新原则 对于众多细胞功能至关重要。