STRUCTURE AND FUNCTION OF THE VACUOLAR ATPASE

液泡ATP酶的结构和功能

• 批准号: 6430865
• 负责人: BARRY BOWMAN
• 金额: $ 28.24万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6430865
• 关键词: Neurospora; active transport; adenosine triphosphate; adenosinetriphosphatase; chemical structure function; electron microscopy; enzyme activity; enzyme structure; gene expression; hydrogen channel; introns; nucleic acid probes; organelles; peptides; protein sequence; proteins; restriction fragment length polymorphism; vesicle /vacuole

The vacuolar ATPase is a large complex enzyme found in the endomembrane system of eucaryotic cells. It functions to acidify the interior of several organelles, e.g., lysosomes and secretory vesicles, and generates an electrochemical gradient to drive transport across the organellar membrane. We have characterized the ATPase in vacuolar membranes from Neurospora and have isolated the genes that encode three of the major subunits. The enzyme appears to have at least six additional subunits. The goals of this proposal are: (1) To isolate the other polypeptides which copurify with ATPase activity, obtain partial amino acid sequence, construct oligonucleotide probes, and isolate the corresponding genes. (2) To use genetics to determine the number of genes essential for synthesizing the vacuolar ATPase. (3) To examine the oligomeric structure of the ATPase by electron microscopy, using antibodies to map the location of several polypeptides. (4) To examine the chromosomal location, exon/intron structure, and the abundance and size of transcripts of vacuolar ATPase genes. Investigation of this enzyme is important because of the central role it plays in the function of many organelles. The uncoupling of ligands and receptors in endosomes, the packaging of neurotransmitters in secretory vesicles, and the hydrolytic action of enzymes in lysosomes all depend on the activity of the vacuolar ATPase. Furthermore, a vacuolar-type ATPase has been reported on the plasma membranes of osteoclasts, where it plays an important role in bone remodeling, and on the plasma membrane of some renal cells, where it plays a role in urinary acidification.

液泡ATPase是一种大型复合酶，发现 桉树细胞系统。 它的功能可以酸化 几个细胞器，例如溶酶体和分泌囊泡，并生成 电化学梯度，可驱动跨行器的运输 膜。 我们已经表征了来自液泡膜的ATPase 神经孢子并隔离了编码三个主要的基因 亚基。 该酶似乎至少具有六个其他亚基。 该建议的目标是：（1）隔离其他多肽 与ATPase活性相关的，获得部分氨基酸序列， 构建寡核苷酸探针，并分离相应的基因。 （2）使用遗传学来确定基因数量 合成液泡ATPase。 （3）检查寡聚 通过电子显微镜的ATPase结构，使用抗体映射 几种多肽的位置。 （4）检查染色体 位置，外显子/内含子结构以及丰度和大小 液泡ATPase基因的转录本。 对该酶的研究很重要，因为它具有核心作用 发挥许多细胞器的功能。 配体的解偶和 内体的受体，分泌中神经递质的包装 囊泡，以及酶在溶酶体中的水解作用都取决于 关于液泡ATPase的活性。 此外，液泡型 已经报道了ATPase在破骨细胞的质膜上 它在骨骼重塑和质膜中起着重要作用 在某些肾细胞中，它在尿酸化中起作用。