Localization of the tight ADP‐binding site on the membrane‐bound chloroplast coupling factor one

Czarnecki, Joseph J.; Abbott, Marilyn S.; Selman, Bruce R.

doi:10.1111/j.1432-1033.1983.tb07699.x

Cited by 26 publications

(9 citation statements)

References 35 publications

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“…8-N 3 -ATP-analogues, on the other hand, most likely adopt a syn conformation in analogy to other nucleotides containing a bulky group at the five-member ring of the purine and thereby representing a conformational change inside the molecule (36,37). A selectivity of incorporation of a 2-N 3 -ATP-analogue in comparison with the 8-N 3 -ATP-analogue has been obtained by nucleotide binding studies with F 1 (38,39) or the fructose 1,6-bisphosphatase (40) with the latter showing a 1000-fold decreased affinity of fructose 1,6-bisphosphatase for the 8-N 3 -ATP-analogue (41) by comparison with the 2-N 3 -ATP-analogue (40). Photoirradiation of F 1 -ATPases with 2-azido-ADP resulted in the exclusive labeling of the ␤ subunit (38,39), thereby discriminating between the catalytic ␤ subunits, where the adenine is in contact with a hydrophobic interface (35), and the noncatalytic ␣ subunits, where binding of adenine involves several hydrogen bonds.…”

Section: Interaction Of the Monofunctional Label 8-n 3 -3ј-biotinyl-amentioning

confidence: 95%

“…A selectivity of incorporation of a 2-N 3 -ATP-analogue in comparison with the 8-N 3 -ATP-analogue has been obtained by nucleotide binding studies with F 1 (38,39) or the fructose 1,6-bisphosphatase (40) with the latter showing a 1000-fold decreased affinity of fructose 1,6-bisphosphatase for the 8-N 3 -ATP-analogue (41) by comparison with the 2-N 3 -ATP-analogue (40). Photoirradiation of F 1 -ATPases with 2-azido-ADP resulted in the exclusive labeling of the ␤ subunit (38,39), thereby discriminating between the catalytic ␤ subunits, where the adenine is in contact with a hydrophobic interface (35), and the noncatalytic ␣ subunits, where binding of adenine involves several hydrogen bonds. This is in contrast to 8-N 3 -ATP or -ADP, which was found to photolabel both the ␣ and ␤ subunits of F 1 (42)(43)(44)(45).…”

Section: Interaction Of the Monofunctional Label 8-n 3 -3ј-biotinyl-amentioning

confidence: 99%

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8-N3-3′-Biotinyl-ATP, a Novel Monofunctional Reagent: Differences in the F1- and V1-ATPases by Means of the ATP Analogue

Schäfer

Coskun

Eger

et al. 2001

Biochemical and Biophysical Research Communications

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Section: Interaction Of the Monofunctional Label 8-n 3 -3ј-biotinyl-amentioning

confidence: 95%

Section: Interaction Of the Monofunctional Label 8-n 3 -3ј-biotinyl-amentioning

confidence: 99%

8-N3-3′-Biotinyl-ATP, a Novel Monofunctional Reagent: Differences in the F1- and V1-ATPases by Means of the ATP Analogue

Schäfer

Coskun

Eger

et al. 2001

Biochemical and Biophysical Research Communications

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“…In particular it is a real substrate (i.e. it is hydrolyzed), and the affinities for the adenine nucleotides and their 2-azido analogues are nearly identical for F-type ATPases (46,47). When TF 0 F 1 was first incubated with 100 M ADP, followed by a second incubation with 100 M 2-N 3 [␣- (Table I, fifth and sixth rows).…”

Section: Discussionmentioning

confidence: 99%

ATP Synthesis by the F0F1-ATPase from the Thermophilic Bacillus PS3 Co-reconstituted with Bacteriorhodopsin into Liposomes

Richard¹,

Pitard²,

Rigaud

1995

Journal of Biological Chemistry

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F-type ATPase from the thermophilic Bacillus PS3, TF 0 F 1 , which was essentially free of bound nucleotides after isolation and purification, was co-reconstituted into liposomes with the light-driven proton pump bacteriorhodopsin. The time course of the light-induced ATP synthesis was biphasic; an initial slow phase accelerated to a final steady-state rate two to three times faster. Adding ATP before initiating the reaction suppressed the slow phase, suggesting that the state of occupancy of specific sites by ATP regulated the synthetic activity of TF 0 F 1 . Incubating the purified TF 0 F 1 with ADP and ATP revealed one ADP and two ATP binding sites that were stable to gel filtration. We analyzed the time courses of light-induced ATP synthesis for the enzyme with different nucleotide content, after co-reconstitution into liposomes with bacteriorhodopsin. The two ATP sites were identified to have regulatory function. A complex containing TF 0 F 1 ⅐ADP, 1:1, was co-reconstituted with various quantities of ATP to obtain a range of molar ratios of TF 0 F 1 ⅐ADP:ATP of between 1:0 and 1:1.7. It was found that the initial rate of ATP synthesis increased with the level of ATP bound to the enzyme. After binding one ATP, a stimulation of ATP synthesis by a factor of 2 was observed. The second ATP site also exhibited regulatory properties. It stimulated ATP synthesis but to a much smaller extent; the stimulation did not exceed 20%. Binding of the photoreactive analogues 2-azido-[␣-32 P]ADP and 2-azido-[␣-32 P]ATP to the TF 0 F 1 and their effects on the rate of ATP synthesis are described further. Importantly, after covalent labeling of the enzyme, tryptic digestion, and high performance liquid chromatography purification, the label was found associated with the ␤Y364-containing tryptic peptide in all cases. ␤Y364 is in the region of conserved residues GXEHYXXA, which is in the ␤ subunit and known to be part of the noncatalytic site.

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“…The tight, energy-dependent ADP binding site on thylakoid-bound CF1 appears to be exclusively located on the/~ subunit as judged by photoaffinity labeling of this site by 2-azido-ADP [36]. The incorporation of analog into this site can be correlated with the inhibition of the ATPase activity of the isolated CF1 [K.R.…”

Section: Chemical Modification and Affinity Labelingmentioning

confidence: 99%

Photosynthetic ATPases: purification, properties, subunit isolation and function

Merchant

Selman

1985

Photosynth Res

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Photosynthetic coupling factor ATPases (F1-ATPases) generally censist of five subunits named α, β, γ, δ and ε in order of decreasing apparent molecular weight. The isolated enzyme has a molecular weight of between 390,000 to 400,000, with the five subunits probably occurring in a 3:3:1:1:1 ratio. Some photosynthetic F1 ATPases are inactive as isolated and require treatment with protease, heat or detergent in order to elicit ATPase activity. This activity is sensitive to inhibition by free divalent cations and appears to be more specific for Ca(2+) vs. Mg(2+) as the metal ion substrate chelate. This preference for Ca(2+) can be explained by the higher inhibition constant for inhibition of ATPase activity by free Ca(2+). Methods for the assay of a Mg-dependent ATPase activity have recently been described. These depend on the presence of organic solvents or detergents in the reaction mixture for assay. The molecular mechanism behind the expression of either the Ca- or Mg-ATPase activities is unknown. F1-ATPases function to couple proton efflux from thylakoid membranes or chromatophores to ATP synthesis. The isolated enzyme may thus also be assayed for the reconstitution of 'coupling activity' to membranes depleted of coupling factor 1.The functions of the five subunits in the complex have been deduced from the results of chemical modification and reconstitution studies. The δ subunit is required for the functional binding of the F1 to the F0. The active site is probably contained in the β (and α) subunit(s). The proposed functions for the γ and ε subunits are, however, still matters of controversy. Coupling factors from a wide variety of species including bacteria, algae, C3 and C4 plants, appear to be immunologically related. The β subunits are the most strongly related, although the α and γ subunits also show significant immunological cross-reactivity. DNA sequence analyses of the genes for the β subunit of CF1 have indicated that the primary sequence of this polypeptide is highly conserved. The genes for the polypeptides of CF1 appear to be located in two cellular compartments. The α, β and ε subunits are coded for on chloroplast DNA, whereas the γ and δ subunits are probably nuclear encoded. Experiments involving protein synthesis by isolated chloroplasts or protein synthesis in the presence of inhibitors specific for one or the other set of ribosomes in the cell suggest the existence of pools of unassembled CF1 subunits. These pools, if they do exist in vivo, probably make up no greater than 1% of the total CF1 content of the cell.

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Localization of the tight ADP‐binding site on the membrane‐bound chloroplast coupling factor one

Cited by 26 publications

References 35 publications

8-N3-3′-Biotinyl-ATP, a Novel Monofunctional Reagent: Differences in the F1- and V1-ATPases by Means of the ATP Analogue

8-N3-3′-Biotinyl-ATP, a Novel Monofunctional Reagent: Differences in the F1- and V1-ATPases by Means of the ATP Analogue

ATP Synthesis by the F0F1-ATPase from the Thermophilic Bacillus PS3 Co-reconstituted with Bacteriorhodopsin into Liposomes

Photosynthetic ATPases: purification, properties, subunit isolation and function

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