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

基本信息

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
项目类别：
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

项目摘要

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）：一个细胞与周围环境的通信对于正常状态和患病状态的众多生理功能至关重要。它要求可以在细胞内“感觉到”与细胞外配体的相遇，相反，可以通过细胞内过程调节细胞外配体 - 受体相互作用的强度。将细胞表面受体的细胞质结构域与细胞骨架（“细胞骨架膜锚”）联系起来的适配器蛋白可能在这种双向通信中起着重要的生物物理作用，即在这种双向交流中起着重要的生物物理作用，即不仅是胞型签名元素，而且是结构性的元素该细胞对机械刺激的反应进行了介导，并可能调节。因此，该应用提出了一个范式，即形式的细胞外配体的串行连接 - 跨膜受体 - 细胞骨架锚定 - 皮质肌动蛋白代表功能单位，这些功能单位是细胞通信中关键生物物理切换板的功能单位。这种序列连接通常会暴露于身体压力，在这种情况下，单个分子相互作用的强度层次结构以及质膜和肌动蛋白皮质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至微观方法。我们建议系统地剖析两种类型的串行跨膜链接的成分的个体贡献。目标1和2将建立涉及整联蛋白LFA-1和E-和N-钙蛋白的串行连接的结构行为。 AIM 1将通过量化各个受体的形成动力和力依赖性失败：配体键（使用尖端的力探测器仪器，即我们的侧视图AFM和光学镊子），将重点放在细胞外结合上。 AIM 2将检查这些受体的细胞骨架锚固的机械行为和分子决定因素。 AIM 3结合力探测和荧光功能成像，将研究该结构组织与细胞函数的相关性。例如，我们将通过机械化学刺激探索白细胞激活的机制。该策略将开辟新的途径，以建立对这些出色的机械信号路径的生物学上合理和物理上现实的理解，从而朝着诊断和治疗疾病的新型自下而上策略。