Regulations and Interactions amoung Molecular Motors

分子马达之间的调节和相互作用

• 批准号: 7691972
• 负责人: YALE E GOLDMAN
• 金额: $ 31.58万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7691972
• 关键词: 3-Dimensional; Actins; Activator Appliances; Address; Adipocytes; Affinity; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Area; Binding; Biochemical; Biochemistry; Biological Assay; Calmodulin; Cell physiology; Cells; Cellular biology; Characteristics; Charcot-Marie-Tooth Disease; Chemistry; Collaborations; Complex; Cytoplasm; Cytoskeleton; Data; Development; Diagnostic; Disease; Dissociation; Dynein ATPase; Electron Microscopy; Environment; Family; Generations; Head; Huntington Disease; Image; In Vitro; Individual; Investigation; Ions; Kinesin; Lateral; Learning; Length; Life; Light; Link; Measures; Mechanics; Microfilaments; Microtubules; Minus End of the Microtubule; Molecular; Molecular Motors; Motion; Motor; Motor Activity; Movement; Myosin ATPase; Myosin Type V; Neck; Neurons; Operative Surgical Procedures; Physiological; Plus End of the Microtubule; Polycystic Kidney Diseases; Power stroke; Principal Investigator; Process; Property; Regulation; Relative (related person); Research; Resolution; Role; Stroke; Structure; System; Techniques; Technology; Total Internal Reflection Fluorescent; Tweens; Upper arm; Vesicle; War; base; behavior observation; cell motility; cellular imaging; dynactin; fundamental research; light microscopy; lissencephaly; motor neuron degeneration; nanometer; optical traps; programs; research study; response; single molecule; structural biology; therapeutic target

The structural changes in unconventional myosins are beginning to be elucidated using a combination of single-molecule mechanical and spectroscopic approaches along with ensemble biochemistry and structural biology. The mechanisms of their regulation and interaction with other molecular motors are still largely unknown. We hypothesize that structural changes in calmodulin molecules bound to IQ motifs in the neck of myosin V and I alter the mechanics of the lever arm when Ca2+ ions bind. Techniques we previously devised for investigation of the basic motility properties will be applied to the mechanism of this modulation. Combined optical trap and single molecule fluorescence microscopy will determine the stiffness changes, rotational motions and rotational mobility of the calmodulin subunits and the relationship of these parameters to modulating motility. Cytoplasmic dynein is a molecular motor that uses the free energy of ATP hydrolysis to drive movement of cargo along microtubules. For a wide range of cellular functions, an activator complex, dynactin, which binds both to dynein and to microtubules is necessary. Dynactin has a role in cargo-binding, but also may also have a more active role in the mechano-chemistry of force generation. Dynein, dynactin, unconventional myosins, and kinesins interact by binding to each other, to individual vesicle cargoes and through their mutual interactions in the cytoplasm. By incorporating several molecular motor types onto manipulatable cargoes in vitro, we will seek to understand these interactions. We will develop assays in vitro that implement aspects of cellular complexity, such as intersections between actin filaments and microtubules near to a surface and away from any surfaces. This work begins a 'bottom up' route to understanding the complexities of cellular motility. We will study the molecular mechanisms of these systems using newly developed technologies, combined optical trap and polarized TIRF microscopy, and 3-D single molecule tracking at nanometer accuracy in vitro and in live cells. These studies complement and link strongly to all of the other sections and cores by providing mechanisms that apply to the cell biological and structural studies with simpler in vitro assemblies of purified cytoskeletal and motor components.

非常规肌凝蛋白的结构变化开始被阐明，使用的组合