Expression Studies of Other Unconventional Myosins

其他非常规肌球蛋白的表达研究

• 批准号: 10699699
• 负责人: James Sellers
• 金额: $ 70.03万
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10699699
• 关键词: Actins; Behavior; Binding; Binding Sites; Biochemical; Biological; Biological Assay; Blindness; Cells; Collaborations; Complement; Drosophila genus; Electrons; Goals; Head; Homologous Gene; Image Analysis; Kinetics; Labyrinth; Length; Mitochondria; Molecular; Motor; Movement; Mutation; Myosin ATPase; Nonmuscle Myosin Type IIA; Pattern; Proteins; Regulation; Retina; Structure; biophysical techniques; deafness; member; microscopic imaging; monomer; optical traps; particle; single molecule

We have begun to analyze the structure and stepping pattern of myosin-6, a processive myosin which moves in the opposite direction on actin compared to other myosins. EM studies show that the angle between the two heads is more variable than in most myosins and that when bound to actin, the motors can be spaced at 13 actins (preferred) or 11 or 15 actin monomers apart. Optical trapping and EM studies demonstrate that the molecule can sometimes take an "inchworm" like step where the two heads occupy closely spaced binding sites. Myosin 7a is present in the stereocilia of the inner ear and in retinal cells. Mutations of this myosin have been associated with deafness and blindness. We have expressed full length mammalian myosin 7a and find that it is autoinhibited, similiar to what we found with its Drosophila homolog. MyoRip, a myosin 7a binding partner, relieves this autoinhibition and allows for processive movements. We are also exploring whether known myosin 7a interacting partners present in the inner ear interact with the myosin and relieve autoinhibition. In collaboration with John Hammer we have begun to study myosin 19 a mitochondrial associated myosin to complement ongoing cell biological studies.

我们已经开始分析肌球蛋白6的结构和步进模式，肌球蛋白6是一种肌球蛋白，与其他肌球蛋白相比，肌动蛋白的方向相反。 EM研究表明，两个头部之间的角度比大多数肌球蛋白更可变，并且在与肌动蛋白界定时，电动机可以在13个肌动蛋白（首选）或11或15个肌动蛋白单体分开。光学诱捕和EM研究表明，该分子有时可以像两个头一样占据紧密间隔的结合位点的“ Inch虫”。 肌球蛋白7a存在于内耳和视网膜细胞的立体膜中。 这种肌球蛋白的突变与耳聋和失明有关。 我们已经表达了全长的哺乳动物肌球蛋白7a，并发现它是自身抑制的，与我们发现的果蝇同源物相似。 肌球蛋白7A绑定伴侣Myorip可以缓解这种自身抑制作用，并允许进行手术运动。 我们还在探索内耳中存在的已知肌球蛋白7a相互作用伴侣是否与肌球蛋白相互作用并缓解自身抑制作用。 通过与约翰·哈默（John Hammer）合作，我们已经开始研究肌球蛋白19 A肌球蛋白相关的肌球蛋白，以补充正在进行的细胞生物学研究。