Diethylstilbestrol. A novel F0-directed probe of the mitochondrial proton ATPase

M. W. McEnery, Peter L Pedersen

Research output: Contribution to journalArticle

Abstract

At low concentrations, diethylstilbestrol (DES) is shown to be a potent F0-directed inhibitor of the F0F1-ATPase of rat liver mitochondria. In analogy to other F0-directed inhibitors, DES inhibits both the ATPase and ATP-dependent proton-translocation activities of the purified and membrane bound enzyme. When added at low concentrations with dicyclohexylcarbodiimide (DCCD), a covalent inhibitor, DES acts synergistically to inhibit ATPase activity of the complex. At higher concentrations, DES restores DCCD-inhibited ATPase activity. However, there is no restoration of ATP-dependent proton translocation. Under these conditions DCCD remains covalently bound to the F0F1-ATPase complex and F1 remains bound to F0. Significantly, when the F0F1-ATPase is inhibited by the F0-directed inhibitor venturicidin rather than DCCD, DES is also able to restore ATPase activity. In contrast, DES is unable to restore ATPase activity to F0F1 preparations inhibited by the F0-directed inhibitors oligomycin or tricyclohexyltin. However, combinations of [DES ± DCCD] or [DES + venturicidin] can restore ATPase activity to F0F1 preparations inhibited by either oligomycin or tricyclohexyltin. Results presented here indicate that the F0 moiety of the rat liver mitochondrial proton ATPase contains a distinct binding site for DES. In addition, they suggest that at saturating concentrations simultaneous occupancy of the DES binding site and sites for either DCCD or venturicidin promote 'uncoupled' ATP hydrolysis.

Original languageEnglish (US)
Pages (from-to)1745-1752
Number of pages8
JournalJournal of Biological Chemistry
Volume261
Issue number4
StatePublished - 1986

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Diethylstilbestrol
Adenosine Triphosphatases
Protons
Dicyclohexylcarbodiimide
Proton-Translocating ATPases
Oligomycins
Adenosine Triphosphate
Liver
Rats
Binding Sites
Mitochondria
Liver Mitochondrion
Restoration
Hydrolysis
Membranes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Diethylstilbestrol. A novel F0-directed probe of the mitochondrial proton ATPase. / McEnery, M. W.; Pedersen, Peter L.

In: Journal of Biological Chemistry, Vol. 261, No. 4, 1986, p. 1745-1752.

Research output: Contribution to journalArticle

McEnery, M. W. ; Pedersen, Peter L. / Diethylstilbestrol. A novel F0-directed probe of the mitochondrial proton ATPase. In: Journal of Biological Chemistry. 1986 ; Vol. 261, No. 4. pp. 1745-1752.
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abstract = "At low concentrations, diethylstilbestrol (DES) is shown to be a potent F0-directed inhibitor of the F0F1-ATPase of rat liver mitochondria. In analogy to other F0-directed inhibitors, DES inhibits both the ATPase and ATP-dependent proton-translocation activities of the purified and membrane bound enzyme. When added at low concentrations with dicyclohexylcarbodiimide (DCCD), a covalent inhibitor, DES acts synergistically to inhibit ATPase activity of the complex. At higher concentrations, DES restores DCCD-inhibited ATPase activity. However, there is no restoration of ATP-dependent proton translocation. Under these conditions DCCD remains covalently bound to the F0F1-ATPase complex and F1 remains bound to F0. Significantly, when the F0F1-ATPase is inhibited by the F0-directed inhibitor venturicidin rather than DCCD, DES is also able to restore ATPase activity. In contrast, DES is unable to restore ATPase activity to F0F1 preparations inhibited by the F0-directed inhibitors oligomycin or tricyclohexyltin. However, combinations of [DES ± DCCD] or [DES + venturicidin] can restore ATPase activity to F0F1 preparations inhibited by either oligomycin or tricyclohexyltin. Results presented here indicate that the F0 moiety of the rat liver mitochondrial proton ATPase contains a distinct binding site for DES. In addition, they suggest that at saturating concentrations simultaneous occupancy of the DES binding site and sites for either DCCD or venturicidin promote 'uncoupled' ATP hydrolysis.",
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