Estimation of Membrane Potential and pH Difference across the Cristae Membrane of Rat Liver Mitochondria

Mitchell, Peter; Moyle, Jennifer

doi:10.1111/j.1432-1033.1969.tb19633.x

Cited by 571 publications

(65 citation statements)

References 27 publications

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“…Movement of protons back into the matrix via the F 1 F O -ATPase then drives phosphorylation of ATP (9,10,20,21) (Figure 11). When electron transport is inhibited, ⌬⌿ m is low, and ATP is available, the F 1 F O -ATPase works in reverse to hydrolyze ATP to extrude protons and restore ⌬⌿ m (16,22,23) (Figure 11).…”

Section: Discussionmentioning

confidence: 99%

F 1 F O-ATPase Activity and ATP Dependence of Mitochondrial Energization in Proximal Tubules after Hypoxia/Reoxygenation

Feldkamp¹,

Kribben²,

Weinberg³

2005

Journal of the American Society of Nephrology

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F reshly isolated proximal tubules subjected to hypoxia/ reoxygenation (H/R) (1) develop a severe mitochondrial functional deficit (1,2) leading to persistent ATP depletion, which plays a pivotal role in overall cellular recovery (3). The energetic deficit occurs despite preserved electron transport (4) and minimal cytochrome c release (2). ATP recovery can be substantially improved by supplementing tubules with citric acid cycle metabolites such as ␣-ketoglutarate and malate individually and in combination (1-3).Mitochondrial membrane potential (⌬⌿ m ), as assessed by 5,5Ј,6,6Ј-tetrachloro-1,1Ј,3,3Ј-tetraethylbenzimidazocarbocyanine iodide (JC-1) uptake is consistently decreased, but not absent, in affected tubules and is restored by the protective citric acid cycle metabolites (1,2,4,5). However, the nonlinearity of formation of JC-1 red aggregates relative to ⌬⌿ m and their slow and incomplete dissociation back to monomers during de-energization limits the use of JC-1 to dynamically follow and manipulate the changes of ⌬⌿ m to gain more mechanistic information about the factors contributing to them (5). Safranin O is concentrated in the mitochondrial matrix as a linear function of ⌬⌿ m where its fluorescence is quenched, allowing its uptake and ⌬⌿ m to be followed by fluorescence changes (6-8). We have recently reported studies employing safranin O in digitonin-permeabilized tubule preparations to alternatively assess ⌬⌿ m in the model (5). Safranin O uptake by mitochondria in digitonin-permeabilized tubules required very small amounts of tubules, permitted measurements of ⌬⌿ m for relatively prolonged periods after the end of the experimental maneuvers, was rapidly reversible during de-energization, and allowed for direct assessment of both substrate-dependent, electron transport-mediated ⌬⌿ m and ATP hydrolysis-supported ⌬⌿ m . Both types of energization in mitochondria of permeabilized tubules measured using safranin O after H/R were impaired. Combining substrates and ATP substantially restored ⌬⌿ m , but did not fully normalize it (5). The objective of the studies reported here was to clarify the mechanism for the energetic deficit by further investigating the role of ATP in facilitating recovery of mitochondrial energization after H/R and determining whether abnormalities of the mitochondrial F 1 F O -ATPase (9,10) or of nucleotide delivery to it by the adenine nucleotide translocase are involved.

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Section: Discussionmentioning

confidence: 99%

F 1 F O-ATPase Activity and ATP Dependence of Mitochondrial Energization in Proximal Tubules after Hypoxia/Reoxygenation

Feldkamp¹,

Kribben²,

Weinberg³

2005

Journal of the American Society of Nephrology

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“…Previous bioenergetic experiments with respiring E. coli and mitochondria have shown that the membrane potential and the rate of respiration are inversely related (5,28). In a regulatory loop known as respiratory control, increases in the membrane potential slow processes that transport protons out of the cell by increasing the energy required to pump protons across the membrane.…”

Section: ϫ2mentioning

confidence: 99%

Enhancement of Survival and Electricity Production in an Engineered Bacterium by Light-Driven Proton Pumping

Johnson

Baron

Naranjo

et al. 2010

Appl Environ Microbiol

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Microorganisms can use complex photosystems or light-dependent proton pumps to generate membrane potential and/or reduce electron carriers to support growth. The discovery that proteorhodopsin is a light-dependent proton pump that can be expressed readily in recombinant bacteria enables development of new strategies to probe microbial physiology and to engineer microbes with new light-driven properties. Here, we describe functional expression of proteorhodopsin and light-induced changes in membrane potential in the bacterium Shewanella oneidensis strain MR-1. We report that there were significant increases in electrical current generation during illumination of electrochemical chambers containing S. oneidensis expressing proteorhodopsin. We present evidence that an engineered strain is able to consume lactate at an increased rate when it is illuminated, which is consistent with the hypothesis that proteorhodopsin activity enhances lactate uptake by increasing the proton motive force. Our results demonstrate that there is coupling of a light-driven process to electricity generation in a nonphotosynthetic engineered bacterium. Expression of proteorhodopsin also preserved the viability of the bacterium under nutrient-limited conditions, providing evidence that fulfillment of basic energy needs of organisms may explain the widespread distribution of proteorhodopsin in marine environments.

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“…In the equation, ⌬⌿ m is the electric potential across the inner mitochondrial membrane, ⌬pH is the pH gradient across the inner membrane, and R, T, and F refer to the gas constant, the absolute temperature, and the Faraday constant, respectively. Under most conditions, ⌬⌿ m is the dominant component of ⌬p, accounting for 150 -180 mV of a ⌬p of 200 -220 mV (107). The ⌬p drives ADP phosphorylation and stops electron flow in the controlled metabolic condition of absence of ADP.…”

Section: Oxidation and Electrochemical Potentials In The Mitochondriamentioning

confidence: 99%

“…The properties of ATP-synthase, called F 1 -ATPase at that time, were described by the work of Racker and co-workers (130) and of Boyer (18). The modern knowledge of the vectorial biochemistry of mitochondrial energy transduction was developed after the formulation of Mitchell's chemiosmotic theory (106,107). The subsequent landmark contribution of Walker (156) was the understanding of the mechanism of ATP synthesis by the molecular rotor driven by proton (H ϩ ) flow through the ATP synthase complex.…”

Section: Biological Energy Transduction: Introduction and Historical mentioning

confidence: 99%

The mitochondrial energy transduction system and the aging process

Navarro¹,

Boveris²

2007

American Journal of Physiology-Cell Physiology

589

401

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Navarro A, Boveris A. The mitochondrial energy transduction system and the aging process. Am J Physiol Cell Physiol 292: C670 -C686, 2007; doi:10.1152/ajpcell.00213.2006.-Aged mammalian tissues show a decreased capacity to produce ATP by oxidative phosphorylation due to dysfunctional mitochondria. The mitochondrial content of rat brain and liver is not reduced in aging and the impairment of mitochondrial function is due to decreased rates of electron transfer by the selectively diminished activities of complexes I and IV. Inner membrane H ϩ impermeability and F1-ATP synthase activity are only slightly affected by aging. Dysfunctional mitochondria in aged rodents are characterized, besides decreased electron transfer and O 2 uptake, by an increased content of oxidation products of phospholipids, proteins and DNA, a decreased membrane potential, and increased size and fragility. Free radicalmediated oxidations are determining factors of mitochondrial dysfunction and turnover, cell apoptosis, tissue function, and lifespan. Inner membrane enzyme activities, such as those of complexes I and IV and mitochondrial nitric oxide synthase, decrease upon aging and afford aging markers. The activities of these three enzymes in mice brain are linearly correlated with neurological performance, as determined by the tightrope and the T-maze tests. The same enzymatic activities correlated positively with mice survival and negatively with the mitochondrial content of lipid and protein oxidation products. Conditions that increase survival, as vitamin E dietary supplementation, caloric restriction, high spontaneous neurological activity, and moderate physical exercise, ameliorate mitochondrial dysfunction in aged brain and liver. The pleiotropic signaling of mitochondrial H 2O2 and nitric oxide diffusion to the cytosol seems modified in aged animals and to contribute to the decreased mitochondrial biogenesis in old animals. oxidative damage; survival; complexes I and IV; nitric oxide synthase EARLY OBSERVATIONS by Leloir and Muñoz (92) indicated that fatty acid oxidation depends on a labile particulate material of liver homogenates. Later, Kennedy and Lehninger (82) reported that the oxidation of fatty acids and of citric acid cycle intermediates was carried out by osmotically active structures with coupled ADP phosphorylation to ATP. Simultaneously, Sjostrand (146) and Palade (126) recognized by electron microscopy the characteristic double membrane of mitochondria. The concept of oxidative phosphorylation as the mitochondrial function evolved from the confluence of structural and biochemical knowledge and by 1952, mitochondria were described by Lehninger (91) as intracellular "power plants".

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Estimation of Membrane Potential and pH Difference across the Cristae Membrane of Rat Liver Mitochondria

Cited by 571 publications

References 27 publications

F 1 F O-ATPase Activity and ATP Dependence of Mitochondrial Energization in Proximal Tubules after Hypoxia/Reoxygenation

F 1 F O-ATPase Activity and ATP Dependence of Mitochondrial Energization in Proximal Tubules after Hypoxia/Reoxygenation

Enhancement of Survival and Electricity Production in an Engineered Bacterium by Light-Driven Proton Pumping

The mitochondrial energy transduction system and the aging process

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