Sequence analysis of the catalytic subunit of H(+)-ATPase from porcine renal brush-border membranes.
The catalytic subunit of the H(+)-ATPase from brush-border membranes of porcine renal proximal tubules was labeled with the hydrophobic SH-group reagent 10-N-(bromoacetyl)amino-1-decyl-beta-glucopyranoside (BADG) which irreversibly inhibits proton pump activity in the absence but not in the presence of ATP. The labeled protein was purified and digested with proteinases. After isolation and sequencing of proteolytic peptides two BADG-labeled cysteines were identified. The amino acid sequences of the obtained proteolytic peptides were homologous to the catalytic subunit of V-ATPases. From mRNA of porcine kidney cortex a catalytic H(+)-ATPase subunit was cloned. 181 of the 183 amino acids which overlap in the sequence derived from the cDNA and the proteolytic peptides were identical, and the two deviations are due to single base exchanges. A comparison of the amino acid sequence derived from the cloned cDNA with sequences of catalytic H(+)-ATPase subunits communicated by other laboratories revealed 98%, 96% and 94% identity with sequences from bovine adrenal medulla, from bovine kidney medulla and from clathrin-coated vesicles of bovine brain. Between 64% and 69% identity was obtained with sequences from fungi and plants. The data show that the catalytic subunit of V-ATPases is highly conserved during evolution. They indicate organ and species specificity in mammalians.
Sander
I
I
Max-Planck-Institut für Biophysik, Frankfurt am Main, Germany.
Lottspeich
F
F
Appelhans
H
H
Kojro
E
E
Spangenberg
J
J
Weindel
C
C
Haase
W
W
Koepsell
H
H
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000737:chemistry; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D004247:DNA; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry; D007672:Kidney Cortex / Q000201:enzymology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008871:Microvilli / Q000201:enzymology / Q000648:ultrastructure; D008969:Molecular Sequence Data; D006180:Proton-Translocating ATPases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013552:Swine
D012333:RNA, Messenger; D004247:DNA; C044848:10-N-(bromoacetyl)amino-1-decylglucopyranoside; D006180:Proton-Translocating ATPases; D000117:Acetylglucosamine