The murine mCLCA3 (alias gob-5) protein is located in the mucin granule membranes of intestinal, respiratory, and uterine goblet cells
The murine mCLCA3 (alias gob-5) protein is located in the mucin granule membranes of intestinal, respiratory, and uterine goblet cells
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DOI：
10.1177/002215540205000609
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发表时间：
2002-06-01
影响因子：
3.2
通讯作者：
Gruber, AD
中科院分区：
文献类型：
--
作者：
Leverkoehne, I;Gruber, AD
推测的阴离子通道mCLCA3(别名GOB-5)是最近发现的钙激活氯离子通道家族(CLCA家族)的第三个成员。初步数据显示，mCLCA3可能在具有分泌功能障碍的疾病中发挥重要作用，包括哮喘和囊性纤维化。本研究对mCLCA3蛋白进行了生化鉴定，并建立了其在正常小鼠组织中的细胞和亚细胞分布模式。制备多克隆兔抗体，并用与mCLCA3多肽的胞外氨基末端相对应的人工合成的寡肽进行亲和免疫纯化。经过体外翻译和糖基化、蛋白酶K保护实验和在COS-7或HEK 293细胞中的异源表达，经SDS-PAGE和免疫印迹分析，显示了与先前鉴定的CLCA蛋白相似的蛋白质结构。一项系统的光、激光共聚焦扫描和透射电子显微镜免疫定位研究，包括几乎所有的小鼠组织，鉴定出mCLCA3蛋白仅与胃肠道、呼吸道和子宫杯状细胞和其他粘蛋白产生细胞的粘蛋白颗粒膜相关。结果提示，mCLCA3可能参与粘蛋白的合成、缩合或分泌。
The putative anion channel mCLCA3 (alias gob-5) is the third murine member of the recently discovered family of calcium-activated chloride channels (CLCA family). Preliminary data suggest that mCLCA3 may play a significant role in diseases with secretory dysfunctions, including asthma and cystic fibrosis. In this study, the mCLCA3 protein was characterized biochemically and its cellular and subcellular distribution pattern was established in normal murine tissues. Polyclonal rabbit antibodies were generated and affinity-immunopurified using synthetic oligopeptides corresponding to the extracellular amino terminus of the mCLCA3 polypeptide. After in vitro translation and glycosylation, proteinase K protection assay, and heterologous expression in COS-7 or HEK 293 cells, SDS-PAGE and immunoblotting revealed a protein structure similar to that of previously characterized CLCA proteins. A systematic light, confocal laser scanning, and transmission electron microscopic immunolocalization study, including virtually all murine tissues, identified the mCLCA3 protein exclusively associated with mucin granule membranes of gastrointestinal, respiratory, and uterine goblet cells and other mucin-producing cells. The results suggest that mCLCA3 may be involved in the synthesis, condensation, or secretion of mucins.