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Thymoquinone Inhibits Escherichia coli ATP Synthase and Cell Growth.

Ahmad Z, Laughlin TF, Kady IO - PLoS ONE (2015)

Bottom Line: The process of inhibition was fully reversible and identical in both membrane bound F1Fo and purified F1 preparations.Moreover, thymoquinone induced inhibition of ATP synthase expressing wild-type E. coli cell growth and non-inhibition of ATPase gene deleted control cells demonstrates that ATP synthase is a molecular target for thymoquinone.This also links the beneficial dietary based antimicrobial and anticancer effects of thymoquinone to its inhibitory action on ATP synthase.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Kirksville College of Osteopathic Medicine, A T Still University of Health Sciences, Kirksville, MO, 63501, United States of America.

ABSTRACT
We examined the thymoquinone induced inhibition of purified F1 or membrane bound F1FO E. coli ATP synthase. Both purified F1 and membrane bound F1FO were completely inhibited by thymoquinone with no residual ATPase activity. The process of inhibition was fully reversible and identical in both membrane bound F1Fo and purified F1 preparations. Moreover, thymoquinone induced inhibition of ATP synthase expressing wild-type E. coli cell growth and non-inhibition of ATPase gene deleted control cells demonstrates that ATP synthase is a molecular target for thymoquinone. This also links the beneficial dietary based antimicrobial and anticancer effects of thymoquinone to its inhibitory action on ATP synthase.

No MeSH data available.


Reversal of TQ induced inhibition by dilution and passing through centrifuge columns.Membrane bound ATP synthase (Mbr) or purified F1 (F1) was inhibited with inhibitory concentration of TQ shown in the figure for 60 min under conditions as described in Fig 2. (A), TrisSO4 pH 8.0 buffer was added to bring back the TQ concentration to non-inhibitory level and activity was measured. (B) Purified F1 was incubated with inhibitory concentrations of TQ for 60 min under conditions as described in Fig 3. Then the inhibited samples were passed twice through 1 ml centrifuge columns and ATPase activity was measured.
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pone.0127802.g004: Reversal of TQ induced inhibition by dilution and passing through centrifuge columns.Membrane bound ATP synthase (Mbr) or purified F1 (F1) was inhibited with inhibitory concentration of TQ shown in the figure for 60 min under conditions as described in Fig 2. (A), TrisSO4 pH 8.0 buffer was added to bring back the TQ concentration to non-inhibitory level and activity was measured. (B) Purified F1 was incubated with inhibitory concentrations of TQ for 60 min under conditions as described in Fig 3. Then the inhibited samples were passed twice through 1 ml centrifuge columns and ATPase activity was measured.

Mentions: TQ induced inhibition of ATP synthase was found to be reversible. Both purified F1 or membranes regained activity after dilution of TQ or removal by passing through centrifuge columns (Fig 4). Again the inhibitory concentrations were determined based on data from Fig 3. The inhibited samples were passed twice through 1 ml centrifuge columns and ATPase activity was measured. It was found that activity was restored to the near normal level as in absence of the TQ (Fig 4). Reversibility data indicates that the observed inhibition is not the result of protein denaturation and that the enzyme retains the ability to reactivate upon release of the compound by dilution or removal through centrifuge columns. Such results indicate non-covalent interaction between TQ and ATP synthase.


Thymoquinone Inhibits Escherichia coli ATP Synthase and Cell Growth.

Ahmad Z, Laughlin TF, Kady IO - PLoS ONE (2015)

Reversal of TQ induced inhibition by dilution and passing through centrifuge columns.Membrane bound ATP synthase (Mbr) or purified F1 (F1) was inhibited with inhibitory concentration of TQ shown in the figure for 60 min under conditions as described in Fig 2. (A), TrisSO4 pH 8.0 buffer was added to bring back the TQ concentration to non-inhibitory level and activity was measured. (B) Purified F1 was incubated with inhibitory concentrations of TQ for 60 min under conditions as described in Fig 3. Then the inhibited samples were passed twice through 1 ml centrifuge columns and ATPase activity was measured.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4440651&req=5

pone.0127802.g004: Reversal of TQ induced inhibition by dilution and passing through centrifuge columns.Membrane bound ATP synthase (Mbr) or purified F1 (F1) was inhibited with inhibitory concentration of TQ shown in the figure for 60 min under conditions as described in Fig 2. (A), TrisSO4 pH 8.0 buffer was added to bring back the TQ concentration to non-inhibitory level and activity was measured. (B) Purified F1 was incubated with inhibitory concentrations of TQ for 60 min under conditions as described in Fig 3. Then the inhibited samples were passed twice through 1 ml centrifuge columns and ATPase activity was measured.
Mentions: TQ induced inhibition of ATP synthase was found to be reversible. Both purified F1 or membranes regained activity after dilution of TQ or removal by passing through centrifuge columns (Fig 4). Again the inhibitory concentrations were determined based on data from Fig 3. The inhibited samples were passed twice through 1 ml centrifuge columns and ATPase activity was measured. It was found that activity was restored to the near normal level as in absence of the TQ (Fig 4). Reversibility data indicates that the observed inhibition is not the result of protein denaturation and that the enzyme retains the ability to reactivate upon release of the compound by dilution or removal through centrifuge columns. Such results indicate non-covalent interaction between TQ and ATP synthase.

Bottom Line: The process of inhibition was fully reversible and identical in both membrane bound F1Fo and purified F1 preparations.Moreover, thymoquinone induced inhibition of ATP synthase expressing wild-type E. coli cell growth and non-inhibition of ATPase gene deleted control cells demonstrates that ATP synthase is a molecular target for thymoquinone.This also links the beneficial dietary based antimicrobial and anticancer effects of thymoquinone to its inhibitory action on ATP synthase.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Kirksville College of Osteopathic Medicine, A T Still University of Health Sciences, Kirksville, MO, 63501, United States of America.

ABSTRACT
We examined the thymoquinone induced inhibition of purified F1 or membrane bound F1FO E. coli ATP synthase. Both purified F1 and membrane bound F1FO were completely inhibited by thymoquinone with no residual ATPase activity. The process of inhibition was fully reversible and identical in both membrane bound F1Fo and purified F1 preparations. Moreover, thymoquinone induced inhibition of ATP synthase expressing wild-type E. coli cell growth and non-inhibition of ATPase gene deleted control cells demonstrates that ATP synthase is a molecular target for thymoquinone. This also links the beneficial dietary based antimicrobial and anticancer effects of thymoquinone to its inhibitory action on ATP synthase.

No MeSH data available.