Primary structure and subunit stoichiometry of F1-ATPase from bovine mitochondria

1H‐NMR studies on nucleotide binding to the catalytic sites of bovine mitochondrial F1‐ATPase

“…We used proton-proton TRNOE measurements to study the nucleotide conformations of various adenine nucleotides bound at these sites. The TRNOE technique involves the extension of classical NOE measurements to exchanging systems such as protein-ligand complexes [8,9], and is ideally suited to study the conformations of ligands bound to very large proteins such as the F1-ATPase (Mr 371 135) [10].…”

The plant mitochondrial F1‐ATPase

“…The purified ATPase migrated as a single protein band on non-denaturing polyacrylamide gel electrophoresis which coincided with the ATPase activity band located by activity staining [15]. Turnip ATPase prepared by this method normally contains six subunits [ [20,21], residues 30-61 of the bovine t5 subunit [18] and residues 3-35 of the sweet potato 8' subunit [12]. Alignments with the sweet potato 8' were carried out manually, those of the chloroplast, E. coli and bovine sequences are taken from Walker et al [18].…”

“…Turnip ATPase prepared by this method normally contains six subunits [ [20,21], residues 30-61 of the bovine t5 subunit [18] and residues 3-35 of the sweet potato 8' subunit [12]. Alignments with the sweet potato 8' were carried out manually, those of the chloroplast, E. coli and bovine sequences are taken from Walker et al [18]. A break of 1 residue was incorporated into the alignments of the chloroplast, E. coli and bovine sequences to maximise homology.…”

“…Fig. 1 shows the 31 residue N-terminal amino acid sequence of turnip Ft-ATPase (5' subunit, manually aligned with sweet potato 8' [12], and with spinach chloroplast CF1 e, bovine 8 and E. coli ~ subunits (aligned by Walker et al [18]). Residue 4 showed evidence of heterogeneity with approximately equal yields of valine and leucine.…”

“…Similarities, albeit weaker, also exist between the turnip ~5 subunit and comparable regions of the bovine F~ ~i and E. coli F~ e subunit N-terminal sequences. Since the chloroplast CFt e, the E. coli ~ and the bovine 8 subunits have been shown to be homologous proteins [18], the turnip F1 8', which shows significant homology with the chloroplast CF1 e subunit, must also be considered homologous to the bovine 8 and E. colie subunits despite the comparatively small number of identical residues in the segment of sequence compared in Fig. 1.…”