High Resolution Characterization of Receptor Signaling and Regulation

受体信号传导和调节的高分辨率表征

• 批准号: 9028523
• 负责人: Barbara A Baird
• 金额: $ 38.12万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-08-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9028523
• 关键词: Actins; Adaptor Signaling Protein; Adherence; Affinity; Allergic; Allergic Disease; Alzheimer' s Disease; Amyloid beta-Protein; Antigens; Area; Basal Cell; Binding; Biological Assay; Biological Models; Brain; Cell Surface Receptors; Cell membrane; Cell model; Cell physiology; Cell surface; Cells; Collaborations; Color; Complex; Computer Simulation; Coupling; Cues; Cytoskeleton; Diffusion; Disease; Endocytosis; Eukaryotic Cell; Event; Extracellular Matrix; F-Actin; Family; Fluorescence Microscopy; Future; Goals; Health; Hydrolysis; IgE; IgE Receptors; Image; Image Analysis; Immune; Immune response; Integrins; Lead; Ligands; Lipids; Measurement; Mechanics; Mediating; Membrane; Membrane Protein Traffic; Microglia; Modeling; Molecular; Mus; NGFR Protein; Neurodegenerative Disorders; Neurologic; Neuronal Plasticity; Neurons; Parkinson Disease; Pattern; Phosphorylation; Phosphotransferases; Play; Process; Property; Proteins; Receptor Signaling; Regulation; Research; Resolution; Role; Signal Pathway; Signal Transduction; Signaling Protein; Spacer DNA; Staging; Stimulus; Surface; Synapses; System; Testing; Time; Translating; alpha synuclein; base; crosslink; extracellular; functional outcomes; inhibitor/antagonist; insight; macrophage; member; nanoscale; neurotrophic factor; prevent; protein complex; receptor; research study; response; sensor

 DESCRIPTION (provided by applicant): This continuing research is aimed at characterizing, with micro- and nanoscale resolution, the collective molecular events of cellular signaling that are initiated by plasma membrane receptors and are highly orchestrated in space and time. Focus remains primarily on the high affinity receptor for IgE, FcεRI, on mast cells, which plays a pivotal role in the allergic immune response and serves as a valuable model system for receptor-mediated cellular signaling. The primary goal of these studies is to elucidate structural interactions of proteins and lipids occurring within the plasma membrane milieu that are altered by antigen crosslinking of IgE- FcεRI and result in transmembrane triggering of intracellular signaling cascades, culminating in cellular responses to the stimulant. Proposed research will investigate mechanisms by which the actin cytoskeleton regulates these interactions at multiple levels, including membrane pinning and compartmentalization by the cortical meshwork that prevents spontaneous initiation of signaling and rearrangements associated with cell adherence to a substrate that adjust the level of response. Specific Aim 1 will use two-color super resolution fluorescence microscopy to image and analyze dynamic interactions among signaling components in the plasma membrane that are initiated immediately after IgE-FcεRI crosslinking, and regulation of these will be evaluated by selective perturbation to membrane and cytoskeleton. Structurally defined ligands will be employed to discern features of crosslinking that are critical for signal initiation, and these experimental results will be incorporated into computational models of signaling systems. Specific Aim 2 will use versatile micro- patterned surfaces and fluorescence microscopy to investigate later-stage membrane interactions initiated by clustered IgE-FcεRI leading to downstream signaling and cellular response events that are modulated by integrin engagement. Selective inhibitors and molecular tension sensors will be employed to probe participation of the actin cytoskeleton and integrins in imposing mechanical forces and regulating signaling networks, as related to consequent cellular responses in single cell assays. Specific Aim 3 will translate biophysical insight and high-resolution approaches gained from detailed characterization of the IgE-FcεRI model system to investigate plasma membrane interactions that participate in neurological cell activities and are disrupted in neurodegenerative diseases. The goal of these highly collaborative studies is to lay the groundwork and establish fruitful directions for significant biomedical impact. Initial experiments will evaluate cellular interactions of α-synuclein that may occur in Parkinson's disease, cell surface clustering of neurotrophin receptor p75 related to signaling in neuronal plasticity, and lysosomal synapses that appear to be formed by microglia to hydrolyze Aβ fibrils associated with Alzheimer's disease.

 描述（由适用提供）：这项持续的研究旨在以微观和纳米级分辨率来表征细胞膜受体启动的细胞信号的集体分子事件，并在时空和时间上进行了高度策划。重点主要保留在肥大细胞上的IgE，FcεRI的高亲和力受体上，该细胞播放A 在过敏性免疫反应中关键作用，并作为受体介导的细胞信号传导的有价值模型系统。这些研究的主要目的是阐明蛋白质和脂质的结构相互作用发生在质膜环境中，这会因Ige-FcεRI的抗原交联改变而改变，并导致跨膜触发细胞内信号级联的跨膜触发，从而使细胞对刺激剂的反应促进。拟议的研究将调查肌动蛋白细胞骨架以多个层次调节这些相互作用的机制，包括通过皮质网固定和隔室化的膜固定和隔室化，从而阻止了信号传导和重新排列与细胞依从性与底物相关的重新排列的启动，从而调整了响应水平。具体的目标1将使用两色超级分辨率荧光显微镜来图像和分析质膜中信号成分之间的动态相互作用，这些信号传导成分在IGE-FCεRI交联后立即启动，并通过选择性扰动与膜和细胞骨骼进行选择性扰动来评估这些相互作用。结构化定义的配体将用于辨别交联的特征，这对于信号启动至关重要，这些实验结果将纳入信号系统的计算模型中。特定的目标2将使用多功能微型表面和荧光显微镜来研究由聚类的IGE-FCεRI引发的后期膜相互作用，从而导致下游信号传导和通过整合素互助调节的细胞反应事件。选择性抑制剂和分子张力传感器将用于探测肌动蛋白细胞骨架和整联蛋白在施加机械力和调节信号网络中的参与，这与随后的单细胞分析中随之而来的细胞反应有关。具体目标3将翻译从IGE-FCεRI模型系统的详细表征中获得的生物物理洞察力和高分辨率方法，以研究参与神经系统细胞活性并在神经退行性疾病中破坏的质膜相互作用。这些高度协作的研究的目的是为重大生物医学影响奠定基础并建立富有成果的方向。初始实验 将评估可能发生在帕金森氏病中的α-突触核蛋白的细胞相互作用，与神经元可塑性信号相关的神经营养蛋白受体p75的细胞表面聚类以及似乎由小胶质细胞形成的溶酶体突触，这些突触似乎是由小胶质细胞形成的。