Valenti G, Verbavatz J M, Sabolić I, Ausiello D A, Verkman A S, Brown D
Renal Unit, Massachusetts General Hospital, Boston.
Am J Physiol. 1994 Sep;267(3 Pt 1):C812-20. doi: 10.1152/ajpcell.1994.267.3.C812.
The water channel CHIP28 accounts for the high water permeability of proximal tubules and thin descending limbs of Henle; a homologous water channel, WCH-CD, in the apical membrane of collecting duct principal cells, may be the vasopressin-sensitive water channel. We show here that one antiserum, raised against CHIP28, immunostains the basolateral membrane of collecting duct principal cells, in addition to staining CHIP28 in other cells. This serum was named anti-basolateral integral protein (anti-BLIP) to distinguish it from other anti-CHIP28 antisera. By Western blotting, BLIP serum recognized both CHIP28 and MIP26, and it stained lens fibers, which contain MIP26 but not CHIP28. BLIP antiserum immunoprecipitated a 28-kDa band, a broad 35- to 50-kDa band, and an approximately 16-kDa band from kidney papilla. It also stained the basolateral membrane of gastric parietal cells, which were not stained with anti-CHIP28 or anti-MIP26 antibodies. BLIP antiserum immunoprecipitated a 28-kDa protein band from stomach; this protein was not precipitated by anti-CHIP28 antibodies. These results suggest that basolateral membranes of principal cells and parietal cells contain a protein(s) that shares common epitopes with CHIP28 and MIP26. Finally, BLIP but not CHIP28 antiserum stained mesothelial (but not epithelial) cells of toad urinary bladder, a further indication that the BLIP antiserum recognizes a protein distinct from CHIP28.
水通道蛋白CHIP28是近端小管和髓袢细降支高水通透性的原因;集合管主细胞顶膜中的一种同源水通道蛋白WCH-CD可能是抗利尿激素敏感的水通道蛋白。我们在此表明,一种针对CHIP28产生的抗血清,除了对其他细胞中的CHIP28进行染色外,还对集合管主细胞的基底外侧膜进行免疫染色。这种血清被命名为抗基底外侧整合蛋白(anti-BLIP),以区别于其他抗CHIP28抗血清。通过蛋白质印迹法,BLIP血清识别CHIP28和MIP26,并且它对含有MIP26但不含有CHIP28的晶状体纤维进行染色。BLIP抗血清从肾乳头免疫沉淀出一条28 kDa的条带、一条宽的35至50 kDa的条带以及一条约16 kDa的条带。它还对胃壁细胞的基底外侧膜进行染色,而抗CHIP28或抗MIP26抗体未对其进行染色。BLIP抗血清从胃中免疫沉淀出一条28 kDa的蛋白条带;这种蛋白不能被抗CHIP28抗体沉淀。这些结果表明,主细胞和壁细胞的基底外侧膜含有一种与CHIP28和MIP26具有共同表位的蛋白质。最后,BLIP抗血清而非CHIP28抗血清对蟾蜍膀胱的间皮(而非上皮)细胞进行染色,这进一步表明BLIP抗血清识别一种不同于CHIP28的蛋白质。