Beta subunit of rat liver mitochondrial ATP synthase: cDNA cloning, amino acid sequence, expression in Escherichia coli, and structural relationship to adenylate kinase

Biochemistry. 1988 Jan 26;27(2):553-60. doi: 10.1021/bi00402a008.

Abstract

The amino acid sequence of all but a few N-terminal residues of the beta subunit of rat liver ATP synthase has been determined from cDNA clones. Rat liver F1-beta is shown to contain 17 amino acid differences from that reported for F1-beta of bovine heart, 2 differences of which involve differences in charge. This may account in part for the observation that bovine heart F1 binds nucleotides with much greater affinity than the rat liver enzyme. Rat liver F1-beta also contains homologous regions with another nucleotide binding protein, adenylate kinase, for which high-resolution structural studies are available. Adjacent to one of these homologous regions is an eight amino acid stretch which bears striking homology to the phosphorylation region of the (Na+,K+)-ATPase. The combination of these two homology regions may constitute at least part of a nucleotide binding domain in F1-beta. Significantly, both rat liver and bovine heart beta contain these regions of homology, whereas the 17 amino acid differences between the two enzymes lie outside this region. The possibility of a second nucleotide binding domain which differs between the two enzymes is discussed. A cDNA clone containing all the regions of homology as well as 11 of the 17 amino acid differences between the bovine heart and rat liver beta subunits has been ligated into the bacterial expression vector pKK223-3. After transformation of a protease-deficient strain of Escherichia coli, this cDNA clone is expressed as a 36-kilodalton protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenylate Kinase / genetics*
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cloning, Molecular*
  • DNA / genetics*
  • Escherichia coli / genetics*
  • Genes
  • Macromolecular Substances
  • Mitochondria, Liver / enzymology*
  • Mitochondrial Proton-Translocating ATPases / genetics*
  • Models, Molecular
  • Molecular Sequence Data
  • Phosphotransferases / genetics*
  • Protein Conformation
  • Proton-Translocating ATPases / genetics*
  • Rats
  • Receptors, Peptide / genetics*
  • Sequence Homology, Nucleic Acid
  • Transcription, Genetic*

Substances

  • ATPase beta-subunit, rat
  • Macromolecular Substances
  • Receptors, Peptide
  • DNA
  • Phosphotransferases
  • Adenylate Kinase
  • Mitochondrial Proton-Translocating ATPases
  • Proton-Translocating ATPases

Associated data

  • GENBANK/M19044