[43] Use of monoclonal antibodies to purify oligomycin sensitivity-conferring protein and to study its interactions with F0 and F1

Archinard, Philippe; Pénin, François; Godinot, Catherine; Gautheron, D

doi:10.1016/s0076-6879(86)26045-0

Cited by 5 publications

(1 citation statement)

References 9 publications

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“…Biological Preparations. Previously described procedures were used to prepare pig heart mitochondria (Gautheron et al, 1964;Godinot et al, 1969), ASU particles (Penin et al, 1986), purified Ft (nucleotide depleted; Penin et al, 1979), purified OSCP (Archinard et al, 1986), and purified protein inhibitor (Di Pietro et al, 1988).…”

Section: Methodsmentioning

confidence: 99%

Fate of nucleotides bound to reconstituted F0-F1 during adenosine 5'-triphosphate synthesis activation or hydrolysis: role of protein inhibitor and hysteretic inhibition

Pénin¹,

Pietro²,

Godinot³

et al. 1988

Biochemistry

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The protein ATPase inhibitor entraps about five nucleotides in pig heart mitochondrial F1, one at least being a triphosphate [Di Pietro, A., Penin, F., Julliard, J.H., Godinot, C., & Gautheron, D.C. (1988) Biochem. Biophys. Res. Commun. 152, 1319-1325]. The fate of these nucleotides was studied during ATP synthesis driven by NADH oxidation in reconstituted inverted submitochondrial particles. Iodinated F1, containing 0.7 mol of endogenous nucleotides/mol, was first loaded with tritiated adenine nucleotides in the presence or absence of the protein inhibitor and then reassociated with F1-depleted submitochondrial particles (ASU particles) to reconstitute an efficient NADH-driven ATP synthesis. In the absence of the protein inhibitor, 1.7 mol of labeled nucleotides remained bound per mole of reassociated F1, 0.8-0.9 mol being rapidly exchangeable against medium ADP or ATP, as measured after rapid filtration through nitrocellulose filters. In the presence of the protein inhibitor, as many as 3.25 mol of labeled nucleotides remained bound per mole of reassociated F1. Under hydrolysis conditions where ATPase activity was highly inhibited, no release of tritiated nucleotide occurred. In contrast, under ATP synthesis conditions where the protonmotive force was generated by NADH oxidation, the progressive reversal of inhibition by the protein inhibitor was correlated to a concomitant release of tritiated nucleotide. When ATP synthesis became fully active, about one nucleotide was completely exchanged whereas more than three nucleotides remained tightly bound and did not appear to be directly involved in ATP synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Methodsmentioning

confidence: 99%

Fate of nucleotides bound to reconstituted F0-F1 during adenosine 5'-triphosphate synthesis activation or hydrolysis: role of protein inhibitor and hysteretic inhibition

Pénin¹,

Pietro²,

Godinot³

et al. 1988

Biochemistry

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Evidence from immunological studies of structure-mechanism relationship of F1 and F1F0

Gautheron¹,

Godinot²

1988

J Bioenerg Biomembr

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Monoclonal and polyclonal antibodies directed against peptides of F1-ATPase of F1F0-ATPase synthase provide new and efficient tools to study structure-function relationships and mechanisms of such complex membrane enzymes. This review summarizes the main results obtained using this approach. Antibodies have permitted the determination of the nature of subunits involved in the complex, their stoichiometry, their organization, neighboring interactions, and vectorial distribution within or on either face of the membrane. Moreover, in a few cases, amino acid sequences exposed on a face of the membrane or buried inside the complex have been identified. Antibodies are very useful for detecting the role of each subunit, especially for those subunits which appear to have no direct involvement in the catalytic mechanism. Concerning the mechanisms, the availability of monoclonal antibodies which inhibit (or activate) ATP hydrolysis or ATP synthesis, which modify nucleotide binding or regulation of activities, which detect specific conformations, etc. brings many new ways of understanding the precise functions. The specific recognition by monoclonal antibodies on the beta subunit of epitopes in the proximity of, or in the catalytic site, gives information on this site. The use of anti-alpha monoclonal antibodies has shown asymmetry of alpha in the complex as already shown for beta. In addition, the involvement of alpha with respect to nucleotide site cooperativity has been detected. Finally, the formation of F1F0-antibody complexes of various masses, seems to exclude the functional rotation of F1 around F0 during catalysis.

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Efficient reconstitution of mitochondrial energy-transfer reactions from depleted membranes and F1-ATPase as a function of the amount of bound oligomycin sensitivity-conferring protein (OSCP)

Pénin

Deléage

Godinot

et al. 1986

Biochimica et Biophysica Acta (BBA) - Bioenergetics

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[43] Use of monoclonal antibodies to purify oligomycin sensitivity-conferring protein and to study its interactions with F0 and F1

Cited by 5 publications

References 9 publications

Fate of nucleotides bound to reconstituted F0-F1 during adenosine 5'-triphosphate synthesis activation or hydrolysis: role of protein inhibitor and hysteretic inhibition

Fate of nucleotides bound to reconstituted F0-F1 during adenosine 5'-triphosphate synthesis activation or hydrolysis: role of protein inhibitor and hysteretic inhibition

Evidence from immunological studies of structure-mechanism relationship of F1 and F1F0

Efficient reconstitution of mitochondrial energy-transfer reactions from depleted membranes and F1-ATPase as a function of the amount of bound oligomycin sensitivity-conferring protein (OSCP)

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