Cytoskeletal Membrane Anchors: Key Switchboards for Cellular Communication, Mecha

细胞骨架膜锚：细胞通信、机甲的关键交换机

• 批准号: 8725191
• 负责人: Volkmar Heinrich
• 金额: $ 29.6万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8725191
• 关键词: Actins; Adhesives; Affinity; Behavior; Binding; Biochemical; Biochemistry; Biomedical Engineering; Cadherins; Calcium; Cell Communication; Cell Surface Receptors; Cell membrane; Cell physiology; Cell surface; Cells; Cellular biology; Chemicals; Chemistry; Collaborations; Communication; Complex; Custom; Cytoplasmic Tail; Cytoskeleton; Defect; Diagnosis; Disease; Dissociation; Docking; Drug usage; Elements; Event; Extracellular Domain; Failure; Fluorescence; Fluorescent Probes; Foundations; Functional Imaging; Future; Generic Drugs; Homing; Immune; Individual; Inflammation; Inflammatory; Integrins; Leukocytes; Life; Ligands; Link; Malignant Neoplasms; Measurement; Measures; Mechanical Stimulation; Mechanical Stress; Mechanics; Mediating; Membrane; Molecular; Molecular Biology; N-Cadherin; Nanotubes; Nature; Neoplasm Metastasis; P-selectin ligand protein; Physiologic pulse; Physiological; Play; Process; Proteins; Reaction; Regulation; Reporter; Research; Role; Rupture; Scheme; Side; Signal Transduction; Site; Spectrum Analysis; Staging; Stimulus; Stress; Structure-Activity Relationship; Techniques; Testing; Time; Translating; Work; adapter protein; base; biophysical tools; cancer cell; cantilever; cell motility; experience; extracellular; fighting; genetic manipulation; innovation; insight; instrument; interest; laser tweezer; leukocyte activation; mechanical behavior; migration; nano; neutrophil; novel; physical property; receptor; receptor binding; research study; response; single molecule; tool

DESCRIPTION (provided by applicant): A cell's communication with its surroundings is central to numerous physiological functions in both normal and diseased states. It requires that encounters with extracellular ligands can be "sensed" inside the cell, and conversely, that the strength of extracellular ligand-receptor interactions can be modulated by intracellular processes. Adapter proteins that link the cytoplasmic domains of cell-surface receptors to the cytoskeleton ("cytoskeletal membrane anchors") are likely to play a crucial biophysical role in this bidirectional communication, i.e., not only as cell-signaling messengers, but also as structural elements that mediate, and possibly regulate, a cell's response to mechanical stimuli. Accordingly, this application puts forward the paradigm that serial linkages of the form extracellular ligand - transmembrane receptor - cytoskeletal anchor - cortical actin represent functional units that act as key biophysical switchboards in cellular communication. Often such serial linkages are exposed to physical stress, in which case the strength hierarchy of individual molecular interactions, along with the physical properties of the plasma membrane and the actin cortex, provide the mechanistic foundation of their mechanoregulatory behavior. Understanding these complex mechanisms requires an interdisciplinary, nano-to-microscale approach. We propose to systematically dissect the individual contributions of the constituents of two types of serial transmembrane linkages. Aims 1 and 2 will establish the structural behavior of serial linkages involving the integrin LFA-1 and E- and N-cadherins. Aim 1 will focus on extracellular binding by quantifying the dynamics of formation and force-dependent failure of the respective receptor:ligand bonds (using cutting-edge force- probe instruments, i.e., our side-view AFM and optical tweezers). Aim 2 will examine the mechanical behavior and the molecular determinants of the cytoskeletal anchors of these receptors. Combining force probing and fluorescent functional imaging, Aim 3 will investigate how this structural organization correlates with cellula function. For example, we will explore the mechanisms of leukocyte activation by mechano-chemical stimuli. This strategy will open new avenues toward establishing a biologically plausible and physically realistic understanding of these remarkable mechanosignaling paths, and thus also toward novel bottom-up strategies for diagnosis and treatment of diseases.

描述（由申请人提供）：细胞与其周围环境的通信对于正常和患病状态下的许多生理功能都至关重要。它要求细胞内可以“感知”与细胞外配体的相遇，相反，细胞外配体-受体相互作用的强度可以通过细胞内过程进行调节。将细胞表面受体的细胞质结构域连接到细胞骨架（“细胞骨架膜锚”）的衔接蛋白可能在这种双向通讯中发挥至关重要的生物物理作用，即不仅作为细胞信号传导信使，而且作为介导并可能调节细胞对机械刺激的反应的结构元件。因此，本申请提出了这样的范例：细胞外配体-跨膜受体-细胞骨架锚定-皮质肌动蛋白形式的串联连接代表在细胞通讯中充当关键生物物理交换机的功能单元。通常，这种串联连接会受到物理压力，在这种情况下，各个分子相互作用的强度等级以及质膜和肌动蛋白皮层的物理特性为其机械调节行为提供了机械基础。了解这些复杂的机制需要跨学科的、纳米到微米尺度的方法。我们建议系统地剖析两种类型的串行跨膜连接成分的个体贡献。目标 1 和 2 将建立涉及整合素 LFA-1 以及 E-和 N-钙粘蛋白的串联连接的结构行为。目标 1 将通过量化相应受体的形成动力学和力依赖性失效来关注细胞外结合：配体键（使用尖端的力探针仪器，即我们的侧视 AFM 和光镊）。目标 2 将检查这些受体的细胞骨架锚的机械行为和分子决定因素。 Aim 3 将力探测和荧光功能成像相结合，研究这种结构组织如何与细胞功能相关。例如，我们将探索机械化学刺激激活白细胞的机制。该策略将为对这些显着的机械信号通路建立生物学上合理且物理上真实的理解开辟新的途径，从而也为疾病诊断和治疗的新的自下而上的策略开辟新的途径。