THE STRUCTURE AND SEQUENCE OF NON-MUSCLE MYOSIN

非肌肉肌球蛋白的结构和序列

• 批准号: 3919993
• 负责人: J A HAMMER
• 金额: --
• 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/3919993
• 关键词: Acanthamoeba; cell biology; cytoskeleton; enzyme structure; genes; genetic library; genetic translation; immunoprecipitation; messenger RNA; molecular biology; molecular cloning; myosins; nucleic acid hybridization; nucleic acid probes; protein structure; site directed mutagenesis

The purpose of this project is to isolate genes encoding the heavy chains of nonmuscle myosins and to use these genes as tools to investigate myosin structure/function relationships and the in vivo functions of nonmuscle myosins. Using molecular cloning techniques, we previously isolated from the soil amoeba Acanthamoeba castellanii the heavy chain genes for myosin II (a nonmuscle myosin possessing conventional structure) and myosin IB (an unusual, low-molecular weight, non-filamentous nonmuscle myosin). In this report we describe the cloning and characterization of two additional amoeba myosin heavy chain genes and our initial efforts at cloning a gene encoding a vertebrate form of myosin I. The significance of this work is that (i) the deduced protein sequences are of great value in furthering our understanding of the structural and functional properties of these myosins and (ii) by using the tools of molecular biology we can approach the study of these proteins in novel ways which are not possible using the classical techniques of protein chemistry. For example, we can dissect at the molecular level the mechanisms of enzymatic regulation using site-directed mutagenesis to alter the myosin molecules and we can examine the physiological roles of these myosins by reintroducing the genes back into cells. Both of these approaches are currently underway.

该项目的目的是隔离编码重型的基因 非肌肉肌球蛋白的链，并将这些基因用作工具 研究肌球蛋白结构/功能关系和体内 非肌肉肌球蛋白的功能。 使用分子克隆 技术，我们以前从土壤变形虫中分离出来 Acanthamoeba Castellanii肌球蛋白II的重链基因（a 具有传统结构的非肌肉肌球蛋白）和肌球蛋白IB （不寻常的，低分子的重量，非丝非肌肉 肌球蛋白）。 在本报告中，我们描述了克隆和 额外的两个变形虫肌球蛋白重链基因的表征 以及我们在克隆编码脊椎动物基因的最初努力 肌球蛋白I的形式。这项工作的意义是（i） 推导的蛋白质序列在推动我们的 了解这些结构和功能特性 肌球蛋白和（ii）通过使用分子生物学工具，我们可以 以新颖的方式对这些蛋白质进行研究 可能使用蛋白质化学的经典技术。 为了 例如，我们可以在分子水平上剖析机制 使用位置定向的诱变来改变酶调节 肌球蛋白分子，我们可以检查 这些肌动物通过将基因重新引入细胞来通过。 两个 这些方法目前正在进行中。