Uncoupling of Oxidative Phosphorylation by Carbonyl Cyanide Phenylhydrazones. II. Effects of Carbonyl Cyanide m-Chlorophenylhydrazone on Mitochondrial Respiration

Goldsby, Richard A.; Heytler, Peter G.

doi:10.1021/bi00905a041

Cited by 52 publications

(29 citation statements)

References 15 publications

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“…Fluorometric measurements were not possible because CCCP fluoresces. Nonetheless, the striking increase in oxygen consumption with the decrease in Jv indicate that CCCP acts under these conditions as a mitochondrial uncoupler similar to its action in other systeins (23). Moreover, the pattern of inhibition of volume, phosphate, and glucose transport caused by CCCP is similar to that observed with arsenate.…”

Section: Resultssupporting

confidence: 55%

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Inhibition of Renal Metabolism. Relative effects of arsenate on sodium, phosphate, and glucose transport by the rabbit proximal tubule.

Brazy¹,

Balaban²,

Gullans³

et al. 1980

J. Clin. Invest.

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A B S T R A C T These studies examine the inhibitory effects of arsenate on the transport of sodium, phosphate, glucose, and para-aminohippurate (PAH) as well as oxidative metabolism by proximal convoluted tubules from the rabbit kidney. Transport rates were measured with radioisotopes in isolated and perfused segments. Metabolic activity was monitored through oxygen-consumption rates and NADH fluorescence in parallel studies in suspensions of cortical tubules. The addition of 1 mM arsenate to the perfusate reduced fluid absorption rates from 1.24±0.17 to 0.66±0.19 nl/mm min (P < 0.01) and lumen-to-bath phosphate transport from 9.93±3.47 to 4.25± 1.08 pmol/mm min (P <0.01). Similar concentrations of arsenate reduced glucose transport only slightly from 66.1±6.0 to 56.8+4.6 pmol/ mm * min (P < 0.05) and had no effect of PAH secretion. Removing phosphate from the perfusate did not affect the net transport of sodium or glucose. In suspensions of tubules, arsenate increased oxygen consunmption rates by 20.5±2.9% and decreased NADH fluorescence by 10.8±1.5)%. These effects on metabolism were concentration dependent and magnified in the presence of ouabain. The data indicate that arsenate's main effect is to uncouple oxidative phosphorylation, and that graded uncoupling of oxidative metabolism causes graded reductions in the net transport of both sodium and phosphate. Glucose transport is inhibited only slightly and PAH secretion is not affected. Thus, partial as opposed to complete inhibition of metabolism reveals that different relationships exist between net Dr. Brazy is a recipient of a Veterans' Administration

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Section: Resultssupporting

confidence: 55%

“…To test this further, we examined the effect of m-chlorocarbonyl cyanide phenylhydrazone (CCCP), a known uncoupler of oxidative metabolism (23). Because of apparent binding of CCCP to serum proteins, these studies were performed using dextran solution as the bathing medium.…”

Section: Resultsmentioning

confidence: 99%

Inhibition of Renal Metabolism. Relative effects of arsenate on sodium, phosphate, and glucose transport by the rabbit proximal tubule.

Brazy¹,

Balaban²,

Gullans³

et al. 1980

J. Clin. Invest.

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“…The concentration of DNP or mii-Cl-CCP which inhibits oxidative phosphorylation is capable of stimulating iboth latent ATPase activity and respiratory rate of mitochondria (8,11,12,19,(21)(22)(23). Furthermore, a current hypothesis is that the primary action of these uncoupling agents is to hydrolyze "highenergy intermediates" which are formed during oxidative phosphorylation (4-7, 19, 20, 22, 23).…”

Section: Resultsmentioning

confidence: 99%

“…The uncoupling concentration of DNP is between 10-5 andl 10-4 m. Recently. Heytler and his collaborators (8,11,12) showed that rni-Cl-CCP was more potent than DXP as an uncoupler of animiial and plant mitochondria. Tight coupling of oxidativ-e phosphorylation is released at rn-CI-CCP concentrations bet-ween 10-and 10-c .…”

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confidence: 99%

Properties of Higher Plant Mitochondria. II. Effects of DNP, m-Cl-CCP, and Oligomycin on Respiration of Mung Bean Mitochondria

Ikuma

Bonner

1967

Plant Physiol.

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Suztenary. Effects of inhlibitors of phosphorvlation on the oxidation of succinate anld of l-malate were investigated with tightl) coupled mitochondria isolated from mung bean hypocotyls. When mitochondria were incubated with 2,4-dinitrophenol, or carbonyl cyanide rn-chloropheilylhydrazone prior to the addition of substrate, the uncoupling effects of these chemicals were relatively small. This is probably caused by relative lack in these mitochondria of endogenous substrates, ATP, and/or "hiigh-energy intermediates". The action of uncoupling agents is, therefore, revealed in a more striking manner when they are introduced during the second state 4. Of the 2 uncoupling agents tested, malate oxidation consistently required 1.5 to 2 times higher concentration of the agents for the half-maximal effects than succinate oxidation. From The classical paper of Loomiiis and Lipmann (21), has been followed by a nunliber of papers showing that DNP3 uncouples phosphorvlation from oxidation in mitochondria isolated both from planlt tissues (1,2,9,10,15,25,26) and from animal tissues (3,4,7, 13,19,23,24). The uncoupling concentration of DNP is between 10-5 andl 10-4 m. Recently. Heytler and his collaborators (8,11,12) showed that rni-Cl-CCP was more potent than DXP as an uncoupler of animiial and plant mitochondria. Tight coupling of oxidativ-e phosphorylation is released at rn-CI-CCP concentrations bet-ween 10-and 10-c . Oligomycin has been shown to inhib.t respiration by blocking a reaction involving the formation of a high energy intelrmlediate (8, 11-13, 16-18, 20, 22, 24 mitochonldria, the oxidationi of NXAD-linked substrates is inhibited sl.ghtly by I jLg of oligomycin, the inhibition is released by D-NP (13,(16)(17)(18).In the literature on plant mitochondrial respiration, emiiphasis hals been placed oIn the similarities between planit and an mal mitochondria (cf. 20). However, our first paper in this series (14) showed that mung bean mitochondria Ppo;sessed several characteristics that were d;fferent from those of animal mitochondr

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“…d) The ratio between the respiratory rate following uncoupling with CCP [31] and the oligomycin inhibited rate (R. C.CCP) [32]. Due to the anomalous response of the system to ADP as discussed below, we did not employ the ratio between the ADP stimulated respiratory rate and the respiratory rate after exhaustion of the ADP supply [33].…”

Section: Respiratory Control Experimentsmentioning

confidence: 99%

Conditions in vitro which Affect Respiratory Control and Capacity for Respiration‐Linked Phosphorylation in Brown Adipose Tissue Mitochondria

Grav

Pedersen

Christiansen

1970

European Journal of Biochemistry

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Oxidative phosphorylation in brown adipose tissue mitochondria is greatly influenced by variations in the in vitro environment. Factors found to be of particular importance were the concentrations of serum albumin, of magnesium and EDTA, and the osmolality and the pH of the medium. The maximal P/O ratios obtained in an isoosmolar medium at pH 6.3, containing 2O//, serum albumin, 7.5 mM MgCl,, and 7.5 mM EDTA with pyruvate-malate as substrate, and using the glucose-hexokinase trapping system, were 2.8 for newborn guinea-pigs and 2.6 for unweaned rats.Under otherwise equal conditions the P/O ratios obtained with 4-week-old guinea-pigs were higher than those with the newborn.EDTA was a requirement for demonstration of respiratory control in brown adipose tissue mitochondria. Respiration with pyruvate-malate or succinate (+ rotenone), and stimulated by inorganic phosphate and ADP, was found to be sensitive to oligomycin and to carbonyl cyanide phenylhydrazone. However, the small extent of stimulation by the phosphate acceptor system, the incomplete inhibition by oligomycin, and the fact that the respiratory rate after exhaustion of added ADP was higher than that prior to the addition of ADP, suggest that brown adipose tissue mitochondria are loosely coupled.The measured respiratory control ratios all increased from the newborn to the 3-week-old guinea-pigs.Mitochondria from newborn and 6-day-old guinea-pigs exhibited the phenomenon of "reversed acceptor control". Thus, in the absence of added inorganic phosphate, ADP inhibited the respiration with pyruvate-malate or with succinate. The reversed ADP control was abolished by carbonyl cyanide phenylhydrazone, but was not suppressed by treatment of the mitochondria with oligomy&.Inorganic phosphate affects respiration in a composite manner. With all of the preparations tested, inorganic phosphate stimulated respiration whether exogenous ADP was present or not.

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Uncoupling of Oxidative Phosphorylation by Carbonyl Cyanide Phenylhydrazones. II. Effects of Carbonyl Cyanide m-Chlorophenylhydrazone on Mitochondrial Respiration

Cited by 52 publications

References 15 publications

Inhibition of Renal Metabolism. Relative effects of arsenate on sodium, phosphate, and glucose transport by the rabbit proximal tubule.

Inhibition of Renal Metabolism. Relative effects of arsenate on sodium, phosphate, and glucose transport by the rabbit proximal tubule.

Properties of Higher Plant Mitochondria. II. Effects of DNP, m-Cl-CCP, and Oligomycin on Respiration of Mung Bean Mitochondria

Conditions in vitro which Affect Respiratory Control and Capacity for Respiration‐Linked Phosphorylation in Brown Adipose Tissue Mitochondria

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