Molecular cloning of cDNA encoding the C subunit of H(+)-ATPase from bovine chromaffin granules.
A cDNA encoding subunit C of the V-ATPase from bovine chromaffin granules was cloned and sequenced. The gene encodes a hydrophilic protein of 382 amino acids with a calculated molecular weight of 43,989. Hydropathy plots revealed no apparent transmembrane segments and a rather high helix content was detected. A cDNA encoding most of the C subunit of the V-ATPase of human brain was also cloned and sequenced. The deduced amino acid sequence of this gene is almost identical to the bovine polypeptide with only one change of tyrosine 336 that was replaced by histidine in the human gene. Two polypeptide fragments derived from subunit E of V-ATPase from chromaffin granules were sequenced and found to be identical to the predicted amino acid sequence of this subunit from bovine kidney. These observations support the idea that the amino acid sequences of corresponding subunits from different V-ATPases are highly conserved. Unlike the A and B subunits of V-ATPases, that are homologous to the beta and alpha subunits of F-ATPases, subunits C and E showed no homology with analogous subunits of the F-ATPase family. It is proposed that the addition of the C and gamma subunits to the respective V- and F-ATPases during evolution defined them as two separate families of H(+)-ATPases.
Nelson
H
H
Roche Institute of Molecular Biology, Roche Research Center, Nutley, New Jersey 07110.
Mandiyan
S
S
Noumi
T
T
Moriyama
Y
Y
Miedel
M C
MC
Nelson
N
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002837:Chromaffin Granules / Q000201:enzymology*; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D015723:Gene Library; D046911:Macromolecular Substances; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000235:genetics*; D012689:Sequence Homology, Nucleic Acid
D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D004247:DNA; D006180:Proton-Translocating ATPases