Properties of Dynein Isoenzymes

动力蛋白同工酶的性质

基本信息

批准号：
7214085
负责人：
IAN R GIBBONS
金额：
$ 28.82万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
1982
资助国家：
美国
起止时间：
1982-04-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7214085
关键词：
ATP Hydrolysis ATP phosphohydrolase Affinity Amino Acid Sequence Amyotrophic Lateral Sclerosis Antibodies Attention Axon Bacteria Basidiomycota Binding Binding Sites Biological Assay Biological Models Biological Process C-terminal Cell division Cell physiology Cilia Class Coiled-Coil Domain Collaborations Complex Coupled Crystallization Cultured Cells Cytoplasm Decompression Sickness Defect Distal Dynein ATPase Electron Microscopy Electrons Epitopes Eukaryota Eukaryotic Cell Face Family Genes Goals Health Human Human Resources Hydrolysis Hylobates Genus In Vitro Indiana Individual Isoenzymes Kinesin Location Maintenance Male Infertility Maps Mechanics Microscopic Microtubules Mitotic spindle Modeling Molecular Conformation Molecular Weight Molecular and Cellular Biology Motor Movement Mutate Mutation Myosin ATPase Nerve Nuclear Nuclear Protein Nuclear Proteins Nucleotides Personal Communication Positioning Attribute Power stroke Principal Investigator Procedures Property Protein Family Protein Isoforms Proteins Protomer Relative (related person)Research Personnel Resolution Roentgen Rays Role Schedule Sequence Analysis Serine-tRNA Ligase Site Site-Directed Mutagenesis Solubility Source Sperm Tail Structure Surface System Techniques Testing Thermus thermophilus Time Transport Vesicles Tubulin Uncertainty Universities Ursidae Family Ustilago Vanadates Yeasts base cell motility cilium motility directed evolution improved in vitro Model member migration molecular mass novel strategies polypeptide programs protein expression research study retrograde transport single molecule size

项目摘要

DESCRIPTION (provided by applicant): Dyneins are motor proteins that move cellular cargos along microtubules toward their minus ends. There are two sub-classes of dynein. Cytoplasmic dynein is a single isoform that is involved in various essential cellular functions, including retrograde vesicle transport in the cytoplasm and in nerve axons, organellar movement, orientation of the mitotic spindle, disassembly of centromeric checkpoint proteins, and nuclear migration. Axonemal dynein occurs as approximately 12 isoforms, encoded by distinct genes, that are responsible for the beating movements of cilia and sperm flagella. In all dyneins the motor unit consists of six AAA protomers disposed in tandem on a single "heavy chain" polypeptide of unusually high molecular mass (>500 kDa). These AAA protomers form a hexameric ring from which the microtubule-binding domain protrudes on a short stalk. Binding and hydrolysis of ATP at certain of the AAA protomers is believed to cause cyclic changes in the angle of the protruding stalk and so translocate the dynein along the microtubule to which it is attached. Sequence analysis suggests that the structure of the AAA protomers in dynein resembles that of other members of the AAA ATPase family, with their closest relationship being to AAA protomers of the recently discovered protein, midasin. The long-term goal of this project is to understand the relationship between structure and function in dynein isoforms. The specific aims are to use techniques of cell and molecular biology: 1) to express individual structural domains of dynein in bacteria or in eukaryote cell cultures in order to obtain relatively low molecular weight forms of these proteins that retain subsets of their functional properties; 2) to use site-directed mutagenesis of the dynein motor to characterize the cooperative ATP-dependent structural changes in the AAA protomers and microtubule-binding stalk that are believed to underlie the ability of dynein to translocate along a microtubule; 3) to use antibodies to define an experimental map of the motor surface and of the changes that occur during the ATPase cycle; 4) to relate the structural and enzymatic properties of the AAA protomers in dynein to those of the AAA protomers in midasin. Defects in dynein underlie many human health problems, including male infertility, bronchiactosis, lissencephaly, primary cilia diskinesia, and defective retrograde transport in nerve axons that can result in amyotrophic lateral sclerosis (ALS).

描述（由申请人提供）：动力蛋白是运动蛋白，它们沿着微管向其减去末端运动。有两个亚基动力蛋白。细胞质动力蛋白是一种参与各种必需细胞功能的单一同工型，包括逆行囊泡在细胞质中和神经轴突中的逆行囊泡转运，细胞器运动，有丝分裂纺锤体的方向，隔离仪表蛋白的分解，隔离。轴突动力蛋白大约是12种同工型，由不同的基因编码，这些基因是纤毛和精子鞭毛的跳动运动。在所有动力蛋白中，运动单元由六个AAA毒剂组成，该毒素在异常高的分子质量（> 500 kDa）的单个“重链”多肽上串联。这些AAA原子体形成一个六聚体环，微管结合结构域在短茎上突出。据信，ATP的结合和水解被认为会导致突出茎的角度的循环变化，因此沿着其附着的微管将动力蛋白转移。序列分析表明，动力蛋白中AAA毒素的结构类似于AAA ATPase家族的其他成员，其最接近的关系是与最近发现的蛋白质Midasin的AAA Frotomers。该项目的长期目标是了解动力蛋白同工型中的结构与功能之间的关系。具体目的是使用细胞和分子生物学技术：1）表达细菌或真核生物细胞培养物中动力蛋白的单个结构结构域，以获得这些蛋白质的相对较低的分子量形式，以保留其功能特性的亚群； 2）使用动力蛋白电动机的位置定向诱变来表征AAA原子剂和微管结合茎的合作ATP依赖性结构变化，这些变化被认为是Dynein沿着微管转化的能力的基础； 3）使用抗体来定义运动表面的实验图和ATPase循环中发生的变化的实验图； 4）将动力蛋白中AAA毒素的结构和酶促特性与Midasin中的AAA frotomer的结构和酶促特性相关联。动力蛋白的缺陷是许多人类健康问题的基础，包括男性不育症，支气管胆汁症，liss脑，原发性纤毛性疾病和神经轴突中有缺陷的逆行转运，这可能导致肌萎缩性侧面硬化症（ALS）。