Biochemical Screens for Modulators of Muscle Force

肌肉力量调节剂的生化筛选

• 批准号: 7532549
• 负责人: Jonathan E. Baker
• 金额: $ 18.55万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7532549
• 关键词: ATP phosphohydrolase; Actin-Binding Protein; Actins; Affect; Arts; Asthma; Binding; Biochemical; Biological Assay; Biophysics; Cells; Development; Disease; Electron Spin Resonance Spectroscopy; Equilibrium; Free Energy; G Actin; Goals; Head; Heart failure; Hypertension; In Vitro; Ionic Strengths; Isometric Exercise; Laboratories; Lasers; Lead; Measurement; Measures; Mechanics; Methods; Modification; Molecular; Muscle; Musculoskeletal; Mutation; Myosin ATPase; Myosin S-1; Numbers; Physiological; Physiology; Protein Isoforms; Proteins; Protocols documentation; Public Health; Range; Sedimentation process; Skeletal Muscle Myosins; Slide; Solutions; Spectrophotometry; Surface; System; Techniques; Temperature; Testing; Theoretical model; Tube; airway hyperresponsiveness; base; body system; cell motility; genetic regulatory protein; high throughput screening; inhibitor/antagonist; novel; novel therapeutics; nucleotide analog; rapid technique; research study; small molecule; small molecule libraries; tool

DESCRIPTION (provided by applicant): Our long term goal is to establish simple biochemical assays that will allow us i) to assess the mechanical effects of a broad range of physiological and disease-related effectors of muscle force, and ii) to rapidly screen small molecule libraries for new therapeutic modulators of muscle force. Our general hypothesis is that most changes in isometric muscle force, F, have a well-defined biochemical basis. Specifically isometric muscle force, F =Funi.N, is influenced by the number, N, of myosin molecules strongly bound to actin and the average force, Funi, per actin bound myosin, where recent studies indicate that Funi varies proportionally with the actin- myosin binding energy, G. The effects of mutations, accessory proteins, and small molecules on the biochemical determinants of isometric muscle force, deltaG and N, can be measured directly in a test tube, suggesting that high throughput screens (HTS) for effectors of muscle force are possible. Using biophysical and biochemical techniques, we have developed several assays that demonstrate the feasibility of a HTS for effectors of muscle force. In this proposal we will (Aim 1) establish a HTS for modulators of muscle force, and we will (Aim 2) verify that these screens are accurate predictors of changes in F, using both in vitro force assays and muscle mechanics experiments. This proposal relies on our laboratory's combined expertise in muscle physiology/biophysics, state-of-the-art biophysical techniques, and theoretical models of muscle mechanochemistry. The HTS for effectors of muscle force developed in this proposal will dramatically accelerate characterization of a wide range of known and suspected modulators of muscle force and will provide a powerful tool for discovering small molecular inhibitors and activators of muscle force. PUBLIC HEALTH RELEVANCE. Fine control of muscle force is central to the normal function, adaptation, and development of most musculoskeletal and organ systems, and disrupting this mechanical balance can lead to a variety of diseases, such as hypertension resulting in cardiac failure and airway hyper-responsiveness associated with asthma. Developing approaches for rapidly assessing the effects of new compounds on muscle mechanics will allow us to rapidly discover new therapies through which we can help muscles in disease states regain their normal mechanical function.

描述（由申请人提供）：我们的长期目标是建立简单的生物化学测定，其将允许我们i）评估广泛的肌肉力量的生理和疾病相关效应物的机械作用，和ii）快速筛选小分子文库以获得肌肉力量的新治疗调节剂。我们的一般假设是，等长肌力F的大多数变化都有明确的生化基础。特别是等长肌力F =Funi.N受与肌动蛋白强结合的肌球蛋白分子的数量N和每个肌动蛋白结合的肌球蛋白的平均力Funi的影响，其中最近的研究表明Funi与肌动蛋白-肌球蛋白结合能G成比例地变化。突变，辅助蛋白，和小分子等长肌力，deltaG和N的生化决定因素的影响，可以直接在试管中测量，这表明高通量筛选（HTS）的肌肉力量的效应是可能的。使用生物物理和生物化学技术，我们已经开发了几种试验，证明了HTS的肌肉力量效应器的可行性。在这个建议中，我们将（目的1）建立一个HTS的肌肉力量的调制器，我们将（目的2）验证这些屏幕是准确的预测F的变化，使用体外力测定和肌肉力学实验。该建议依赖于我们实验室在肌肉生理学/生物物理学，最先进的生物物理技术和肌肉机械化学理论模型方面的综合专业知识。在这项建议中开发的肌肉力量效应器的HTS将大大加快表征广泛的已知和可疑的肌肉力量调节剂，并将为发现肌肉力量的小分子抑制剂和激活剂提供一个强大的工具。公共卫生相关性。肌肉力量的精细控制对于大多数肌肉骨骼和器官系统的正常功能、适应和发育至关重要，破坏这种机械平衡可能导致各种疾病，例如导致心力衰竭的高血压和与哮喘相关的气道高反应性。开发快速评估新化合物对肌肉力学影响的方法将使我们能够快速发现新的疗法，通过这些疗法，我们可以帮助处于疾病状态的肌肉恢复正常的机械功能。