STRUCTURE AND FUNCTION OF DYNEIN MOLECULES

动力蛋白分子与纤毛和鞭毛微管相互作用， 产生纤毛搏动，伴随着ATP的水解。 一个 了解它们的作用方式可能有助于分析这些问题。 临床疾病如不孕症、肺气肿和囊性纤维化， 属于有丝分裂纺锤体的力学和微管介导的 在胚胎发育过程中形状发生变化。 方法已经在这些方面得到了发展。 分离和纯化动力蛋白的实验室 和内臂，并在原位和体外可视化这些分子， 使用速冻深蚀刻技术。 长期目标是 以现在可能的方式来理解动力蛋白力产生的循环 对于肌球蛋白循环。 五个具体目标被描述。 1.外臂 动力蛋白将被解离成亚单位， 高效液相色谱法 通过将深蚀刻与 ATP酶活性，微管结合能力， 单克隆抗体结合等，应该可以 生成蛋白质结构域的功能图。 2.外臂动力蛋白 将被研究，因为它装饰微管和响应ATP，目标是 分析其ATP和微管蛋白的定位和修饰 结合位点。 这些研究有望补充关于 解离动力蛋白 3.内臂动力蛋白的功能图将是 产生，并在睫状体力学的作用将进行评估。 4. 来自各种来源的不寻常的动力蛋白或动力蛋白样蛋白将被 研究以建立同源性，并寻找功能相似性， “标准”动力蛋白。 5.将研究洗涤剂提取的运动细胞， 了解动力蛋白如何改变其构型， 磷酸盐协会，在原位跳动周期。

Dynein molecules interact with ciliary and flagellar microtubules to generate the ciliary beat, with the concomitant hydrolysis of ATP. An understanding of their mode of action may facilitate the analysis of such clinical disorders as infertility, emphysema, and cystic fibrosis, and may pertain to the mechanics of the mitotic spindle and to microtubule-mediated shape changes during embryology. Methods have been developed in these laboratories for the isolation and purification of dyneins from the outer and inner arms, and for visualizing these molecules in situ and in vitro using the quick-freeze deep-etch technique. The long-term objective is to understand the dynein force-generating cycle in the fashion now possible for the myosin cycle. Five specific aims are described. 1. Outer-arm dynein will be dissociated into subunits and fractionated by high-performance liquid chromatography. By combining deep-etching with studies on ATPase activity, microtubule-binding ability, monoclonal-antibody binding, and the like, it should be possible to generate a functional map of the protein's domains. 2. Outer-arm dynein will be studied as it decorates microtubules and responds to ATP, the goal being to analyze the location and the modification of its ATP and tubulin binding sites. These studies will hopefully complement the studies on dissociated dynein. 3. A functional map of the inner-arm dyneins will be generated, and their role in ciliary mechanics will be assessed. 4. Unusual dyneins or dynein-like proteins from various sources will be studied to establish homologies and look for functional similarities with "standard" dyneins. 5. Motile detergent-extracted cells will be studied to learn how dyneins change their configuration, and their nucleotide phosphate associations, during the in situ beat cycle.