Deciphering the functional role of actin-spectrin-based membrane skeleton in subcellular compartmentalization of signaling proteins and cell signal transduction

破译基于肌动蛋白-血影蛋白的膜骨架在信号蛋白的亚细胞区室化和细胞信号转导中的功能作用

• 批准号: 10580148
• 负责人: Ruobo Zhou
• 金额: $ 24.98万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10580148
• 关键词: Actins; Affect; Behavior; Binding; Biochemical; Cell Adhesion Molecules; Cell Communication; Cell Differentiation process; Cell Proliferation; Cell Surface Proteins; Cell Survival; Cell membrane; Cell surface; Cells; Cellular biology; Clinical; Complex; Development; Disease; Drug Targeting; Endocytosis; Eukaryotic Cell; Family; G-Protein-Coupled Receptors; Goals; Human; Image; Intracellular Membranes; Liquid substance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Membrane; Membrane Proteins; Metabolism; Molecular; Molecular Biology; Mutation; Neurodegenerative Disorders; Neurons; Organelles; Outcome; Periodicity; Phase; Protein Family; Proteins; Proteome; Receptor Protein-Tyrosine Kinases; Research; Resolution; Role; Sensitivity and Specificity; Signal Pathway; Signal Transduction; Signaling Protein; Site; Skeleton; Spectrin; Stimulus; Structure; Syndrome; Therapeutic Intervention; Transactivation; base; biophysical properties; extracellular; first responder; human disease; insight; membrane skeleton; nanocluster; new therapeutic target; novel; novel therapeutic intervention; protein complex; receptor; spatiotemporal; targeted treatment; therapy design; tool

Project Summary/Abstract: Receptor tyrosine kinases (RTKs), G-protein-coupled receptors (GPCRs), and cell adhesion molecules (CAMs) are three major families of cell surface proteins in all eukaryotic cells, and together represent the primary first responders for cells to respond an extracellular stimulus and initiates a variety of signaling pathways to subsequently regulate cell proliferation and differentiation, promote cell survival, and modulate cellular metabolism and cell-to-cell communication. Mutations affecting these signaling pathways result in many human syndromes and diseases, such as various types of neurodegenerative disorders and cancer. The clinical importance of these signaling proteins has motivated the development of targeted therapies designed to block the activation of the membrane receptors and the downstream signal transduction. Increasing evidence has suggested that there is significant signaling crosstalk between these three membrane protein families at the plasma membrane level and these proteins can form highly organized membrane micro- or nano-clusters with unique biochemical and biophysical properties, dictating the signaling outcome. However, the molecular mechanisms by which how such crosstalk and compartmentalization of membrane-associated signaling proteins are initiated to modulate the sensitivity and specificity of the downstream signaling remain largely elusive. Our recent discovery of a newly identified actin-spectrin-based membrane-associated periodic skeleton (MPS) structure being a signaling platform for RTK transactivation by GPCRs and CAMs in neurons provides molecular insights into how the cooperative action among these cell surface proteins can be coordinated to give rise to the downstream signaling. The objective of this proposal is to combine super-resolution imaging, cell and molecular biology tools, and mass spectrometry analyses, to investigate the detailed molecular mechanisms responsible for the MPS-mediated cell signaling, by identifying the key molecular interactions responsible for the MPS-dependent initiation of the signaling protein clustering (i.e., complex formation) and examining the roles of liquid-liquid phase separation, receptor endocytosis, and contact sites between the plasma membrane and intracellular membrane-bound intracellular organelles in the MPS-mediated signaling. Our proposed research will not only broaden our fundamental understanding of cell signal transduction controlled by the membrane skeleton and the phase separation behaviors of signaling proteins, but also help suggest new drug targets for human diseases including neurodegenerative diseases and cancer.

项目摘要/摘要： 受体酪氨酸激酶（RTK），G蛋白偶联受体（GPCR）和细胞粘附 分子（CAM）是所有真核细胞中细胞表面蛋白的三个主要家族，并且 共同代表细胞反应细胞外刺激的主要第一反应者 启动各种信号通路，以调节细胞增殖和 分化，促进细胞存活，并调节细胞代谢和细胞对细胞的新代谢 沟通。影响这些信号通路的突变导致许多人类综合症 和疾病，例如各种类型的神经退行性疾病和癌症。临床 这些信号蛋白的重要性促使有针对性疗法的发展 设计用于阻断膜受体的激活和下游信号 转导。越来越多的证据表明存在明显的信号串扰 在质膜水平的这三个膜蛋白家族和这些蛋白之间 可以形成具有独特的生化和 生物物理特性，决定了信号传导结果。但是，分子机制通过 这种串扰和与膜相关信号蛋白的分隔如何 开始调节下游信号的灵敏度和特异性，在很大程度上保持不变 难以捉摸。我们最近发现了新确定的基于肌动蛋白的膜相关的 周期性骨骼（MPS）结构是GPCR进行RTK反式激活的信号平台 神经元中的凸轮提供了分子见解，以了解它们之间的合作作用如何 细胞表面蛋白可以协调以产生下游信号传导。目标 该建议的是结合超分辨率成像，细胞和分子生物学工具，以及 质谱分析，以研究负责的详细分子机制 MPS介导的细胞信号传导，通过识别负责的关键分子相互作用 信号蛋白聚类的MPS依赖性启动（即复合形成）和 检查液 - 液相分离，受体内吞作用和接触部位的作用 在质膜和细胞内膜结合的细胞内细胞器之间 MPS介导的信号传导。我们提出的研究不仅会扩大我们的基本 了解由膜骨骼控制的细胞信号转导和相位 信号蛋白的分离行为，但也有助于暗示人类的新药物目标 包括神经退行性疾病和癌症在内的疾病。