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GASTRIC ACID SECRETION: RECONSTITUTION OF PROTON PUMP

胃酸分泌：质子泵的重建

基本信息

批准号：
3153124
负责人：
EDWIN C RABON
金额：
$ 10.98万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
1984
资助国家：
美国
起止时间：
1984-08-01 至 1987-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3153124
关键词：
adenosinetriphosphatase gastric acid gastric juice liposomes phospholipids potassium chloride secretion

项目摘要

The long term goal of this poject is to determine the mechanisms responsible for H+ and Cl- secretion in the stomach. It is known that H+ secretion is dependent upon H+/K+ exchange activity of the (H+,K+) ATPase and that in the non-secreting state this activity is limited by K+ accessibility at a K+ dependent catalytic site located within the tubulovesicles of the parietal cell. Following the morophological changes accompanying the transition from resting to secreting state this ATPase is localized within the intracellular canaliculus. We will investigate the pump complex isolated from either resting or stimulated tissue to determine what changes this morphological transformation exerts on the basic mode of pump operation and ancillary routes of K+ and Cl- permeability. In the short term, this proposal addresses the solubilization, purification and reconstitution of the gastric proton transport system. The detergent, n-octylglucoside, will be utilized to extract the active solubilized (H+,K+) ATPase from both resting and stimulated mucosa. Immunoaffinity chromatography using the monoclonal antibody H,K III will then be used to purify the solubilized, active enzyme. This purified protein will then be inserted into artificial phospholipid vesicles and studied by optical probes of pH and membrane potential. A possible significance of this work is in the treatment of ulcer disease. Inhibition of this enzyme by the substituted benzimidazoles has been correlated with inhibition of acid secretion in humans. A detailed knowledge of this ATPase is necessary to design inhibitors of acid secretion which are selective for this enzyme.

本项目的长期目标是确定负责胃中的H+和Cl-分泌。已知H+ 分泌依赖于（H+，K+）ATP酶的H+/K+交换活性在非分泌状态下，这种活性受到K+的限制可及性在K+依赖的催化位点位于壁细胞的管状小泡。随着形态学的变化伴随着从静息状态到分泌状态的转变，该ATP酶位于细胞内小管内。我们将调查从静止或刺激的组织中分离的泵复合物，以确定是什么改变了这种形态转换对基本模式的影响，泵操作和K+和Cl-渗透率的辅助路线。在短期内，该建议解决了溶解、纯化和胃质子转运系统的重建。清洁剂，正辛基葡糖苷，将用于提取活性溶解的（H+，K+）ATP酶从静止和刺激粘膜。免疫亲和使用单克隆抗体H，K III的层析然后将用于纯化溶解的活性酶。然后将纯化的蛋白质插入到人造磷脂囊泡中， pH和膜电位的探针。这项工作的一个可能的意义是用于治疗溃疡病这种酶的抑制作用，取代的苯并咪唑与酸的抑制有关人类的分泌物。有必要详细了解这种ATP酶，设计对这种酶有选择性的酸分泌抑制剂。