title: The epithelial sodium channel. Subunit number and location of the amiloride binding site.
abstract: Sodium dodecyl sulfate gel electrophoresis of the radioiodinated native amiloride-sensitive epithelial sodium channel protein isolated from bovine renal papilla and cultured amphibian A6 cells under denatured and nonreduced conditions revealed an 125I-labeled protein band of Mr approximately 730,000. Upon reduction, this protein was resolved into five major polypeptide bands with apparent average Mr values of 315,000, 149,000, 95,000, 71,000, and 55,000. The amiloride analog [3H]methylbromoamiloride has been used as a photoaffinity label to determine the location of the binding site for amiloride on the epithelial sodium channel protein. [3H]Methylbromoamiloride binds covalently to the sodium channel at high affinity binding sites with a half-maximal binding concentration of 0.2 microM. [3H]Methylbromoamiloride was specifically photoincorporated into the Mr approximately 150,000 polypeptide and this incorporation was blocked by addition of excess amiloride. These data suggest that the epithelial sodium channel protein is composed of at least five nonidentical polypeptide subunits, only one of which specifically binds amiloride.
mesh_terms: D000345:Affinity Labels; D000584:Amiloride / Q000378:metabolism*; D000663:Amphibians; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002460:Cell Line; D004848:Epithelium / Q000032:analysis; D007473:Ion Channels / Q000378:metabolism*; D007668:Kidney / Q000032:analysis*; D008970:Molecular Weight; D010777:Photochemistry; D012964:Sodium / Q000378:metabolism*
chemical_list: D000345:Affinity Labels; D007473:Ion Channels; D000584:Amiloride; D012964:Sodium