Primary sequence and functional expression of a novel beta subunit of the P-ATPase gene family.
The cortical collecting tubule (CCT) of the mammalian kidney reabsorbs sodium and potassium, processes that are mediated by Na/K-ATPase and H/K-ATPase. CCT is also an important site for proton secretion, which is driven, in part, by H/K-ATPase. Na/K-ATPase and H/K-ATPase are members of the ion-motive P-ATPase gene family. They are closely related plasma membrane proteins which consist of alpha beta heterodimers. The urinary bladder of the toad Bufo marinus is the amphibian counterpart of mammalian CCT. We have previously characterized a ouabain-resistant Na/K-ATPase [see ref. 17], from TBM cells, a clonal cell line derived from the toad bladder, which expresses transepithelial sodium transport. In the present study, we report the primary sequence and functional expression of a novel beta subunit (beta bladder = beta b1) isolated from a toad bladder epithelial cell cDNA library. The deduced polypeptide is 299 amino acids in length and has a predicted molecular mass of 33 kDa. The beta b1 protein exhibits 35% amino acid identity to the previously characterized beta 1 of B. marinus Na/K-ATPase and 39% identity with beta 3 of B. marinus Na/K-ATPase. It shares 38% identity with the mammalian beta gastric H/K-ATPase and 52% with the mammalian beta 2 Na/K-ATPase. Northern blot analysis shows that a 1.4 x 10(3)-base mRNA is expressed at a high level in bladder epithelial cells and eye and at a trace level in kidney; it is not detectable in significant amounts in the stomach, colon and small intestine.(ABSTRACT TRUNCATED AT 250 WORDS)
Jaisser
F
F
Institut de Pharmacologie et de Toxicologie de l'Université, Lausanne, Switzerland.
Horisberger
J D
JD
Rossier
B C
BC
D000251:Adenosine Triphosphatases / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002024:Bufo marinus / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D014982:Xenopus laevis
D018076:DNA, Complementary; D012333:RNA, Messenger; D000251:Adenosine Triphosphatases; D000254:Sodium-Potassium-Exchanging ATPase