Distribution and diversity of Na-K-Cl cotransport proteins: a study with monoclonal antibodies

• DOI: 10.1152/ajpcell.1995.269.6.c1496
• Publication Date: 2017-12-24T19:22:16Z
• Authors: C. Lytle; J. C. Xu; D. Biemesderfer; B. Forbush
• Keywords: Immunoprecipitation

The Na-K-Cl cotransporter (NKCC) is present in most animal cells where it functions cell volume homeostasis and epithelial salt transport. We developed six monoclonal antibodies (designated T4, T8, T9, T10, T12, T14) against a fusion protein fragment encompassing the carboxy-terminal 310 amino acids of human colonic NKCC. These T selectively recognized putative NKCC proteins diverse variety tissues. Western blot analysis membranes isolated from 23 types identified single bands immunoreactive ranging mass 146 to 205 kDa. amount detected these correlated with loop diuretic binding site density. Proteins previously as cotransporters by photoaffinity labeling were mutually multiple antibodies. Most effectively immunoprecipitated denatured form protein. Immunocytochemical studies on rabbit parotid gland demonstrated that restricted basolateral margin acinar absent ducts, accord central role cotransport chloride secretion. In kidney, was localized apical membrane thick ascending limb cells, consistent its reabsorption.