Gastric parietal cell H(+)-K(+)-ATPase microsomes are associated with isoforms of ankyrin and spectrin.

Gastric parietal cell H(+)-K(+)-ATPase microsomes are associated with isoforms of ankyrin and spectrin.
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胃壁细胞 H( )-K( )-ATP 酶微粒体与锚蛋白和血影蛋白同工型相关。

DOI:
10.1152/ajpcell.1993.264.1.c63
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发表时间:
1993
期刊:
The American journal of physiology
影响因子:
--
通讯作者:
Saccomani,G
Saccomani,G
中科院分区:
--
文献类型:
--
作者:
Smith,PR;Bradford,AL;Joe,EH;Angelides,KJ;Benos,DJ;Saccomani,G

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胃壁细胞刺激HCl分泌需要细胞质H(+)-K(+)- atp酶的小管囊泡与顶膜融合。膜的插入通过微绒毛的形成导致根尖表面积的急剧增加。为了阐明可能稳定顶端膜内新插入的H(+)-K(+)- atp酶的元件,我们寻找了与胃酶相关的特定细胞骨架蛋白。我们通过免疫印迹分析证明,锚蛋白、谱蛋白和肌动蛋白与从胃壁细胞制备的H(+)-K(+)- atp酶微粒体共化。使用抗H(+)-K(+)- atp酶抗体将125i标记的天然红细胞锚蛋白与来自胃微粒体的H(+)-K(+)- atp酶共沉淀,表明锚蛋白与H(+)-K(+)- atp酶相关。间接免疫荧光和共聚焦显微镜显示,锚蛋白和H(+)-K(+)- atp酶在静息和分泌的壁细胞内共分离。综上所述,这些数据表明,胃H(+)-K(+)- atp酶与谱蛋白和肌动蛋白的关联是由锚蛋白介导的,这种相互作用有助于在胃酸分泌过程中维持酶在胃壁细胞顶域的极化分布。
Stimulation of HCl secretion by gastric parietal cells requires the fusion of cytoplasmic H(+)-K(+)-ATPase-bearing tubulovesicles with the apical membrane. This insertion of membrane results in a dramatic increase in apical surface area through the formation of microvilli. To elucidate the elements that may stabilize the newly inserted H(+)-K(+)-ATPase within the apical membrane, we searched for specific cytoskeletal proteins associating with the gastric enzyme. We document by immunoblot analysis that ankyrin, spectrin, and actin copurify with H(+)-K(+)-ATPase microsomes prepared from gastric parietal cells. Coprecipitation of 125I-labeled native erythrocyte ankyrin with the H(+)-K(+)-ATPase from gastric microsomes using anti-H(+)-K(+)-ATPase antibodies suggests that ankyrin associates with the H(+)-K(+)-ATPase. Indirect immunofluorescence and confocal microscopy show that ankyrin and H(+)-K(+)-ATPase cosegregate within resting and secreting parietal cells. Taken together, these data suggest that the association of the gastric H(+)-K(+)-ATPase with spectrin and actin is mediated by ankyrin and that this interaction contributes to the maintenance of the polarized distribution of the enzyme to the apical domain of gastric parietal cells during acid secretion.
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