Gnaiger 1995 J Bioenerg Biomembr

Gnaiger E, Steinlechner-Maran R, Méndez G, Eberl T, Margreiter R (1995) Control of mitochondrial and cellular respiration by oxygen. J Bioenerg Biomembr 27:583-96.

» PMID: 8746845

Gnaiger E, Steinlechner-Maran R, Mendez G, Eberl T, Margreiter R (1995) J Bioenerg Biomembr

Abstract: Control and regulation of mitochondrial and cellular respiration by oxygen is discussed with three aims: (1) A review of intracellular oxygen levels and gradients, particularly in heart, emphasizes the dominance of extracellular oxygen gradients. Intracellular oxygen pressure, p_O2, is low, typically one to two orders of magnitude below incubation conditions used routinely for the study of respiratory control in isolated mitochondria. The p_O2 range of respiratory control by oxygen overlaps with cellular oxygen profiles, indicating the significance of p_O2 in actual metabolic regulation. (2) A methodologically detailed discussion of high-resolution respirometry is necessary for the controversial topic of respiratory control by oxygen, since the risk of methodological artefact is closely connected with far-reaching theoretical implications. Instrumental and analytical errors may mask effects of energetic state and partially explain the divergent views on the regulatory role of intracellular p_O2 . Oxygen pressure for half-maximum respiration, p₅₀, in isolated mitochondria in the LEAK state was 0.025 kPa (0.2 Torr; 0.3 µM O₂), whereas p₅₀ in endothelial cells was 0.06–0.08 kPa (0.5 Torr). (3) A model derived from the thermodynamics of irreversible processes was developed which quantitatively accounts for near-hyperbolic flux/p_O2 relations in isolated mitochondria. The apparent p₅₀ is a function of redox potential and protonmotive force. The protonmotive force collapses after uncoupling and consequently causes a decrease in p₅₀. Whereas it is becoming accepted that flux control is shared by several enzymes, insufficient attention is paid to the notion of complementary kinetic and thermodynamic flux control mechanisms. • Keywords: Oxygen limitation, p₅₀, Critical oxygen pressure, Respirometry, Polarographic oxygen sensor, Human endothelial cells, Rat liver mitochondria, Intracellular pO₂, Oxygen gradients, Kinetics, Nonequilibrium thermodynamics

• O2k-Network Lab: AT_Innsbruck_Gnaiger E

Labels: MiParea: Respiration, Instruments;methods 

Stress:Hypoxia  Organism: Human, Rat  Tissue;cell: Liver, Endothelial; Epithelial; Mesothelial Cell"Endothelial; Epithelial; Mesothelial Cell" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property.  Preparation: Intact cells, Isolated Mitochondria"Isolated Mitochondria" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property. 

Regulation: ADP, Flux control, O2"O2" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.  Coupling state: LEAK, ROUTINE, OXPHOS 

HRR: Oxygraph-2k 

Made history 

High-resolution respirometry

• This publication was the first to establish high-resolution respirometry, based on the OROBOROS Oxygraph [1] and integrating with DatLab [2,3] the standardized oxygen calibration (air and internal zero oxygen calibration), automatic instrumental background correction of oxygen flux, deconvolution of the oxygen signal for time resolution required in oxygen kinetics and quality control of the limit of detection of oxygen flux at ±1 pmol O₂·s^-1·ml^-1.

1. Haller T, Ortner M, Gnaiger E (1994) A respirometer for investigating oxidative cell metabolism. Toward optimization of respiratory studies. Analyt Biochem 218:338-42. »PMID: 8074290
2. Méndez G, Gnaiger E (1994) How does oxygen pressure control oxygen flux in isolated mitochondria? A methodological approach by high-resolution respirometry and digital data analysis. In: What is Controlling Life? (Gnaiger E, Gellerich FN, Wyss M, eds) Modern Trends in BioThermo-Kinetics 3. Innsbruck Univ Press:191-4. »Open Access
3. Steinlechner-Maran R, Eberl T, Margreiter R, Gnaiger E (1994) Oxygen dependence of cellular respiration in endothelial cells: A sensitive toxicological test. In: What is Controlling Life? (Gnaiger E, Gellerich FN, Wyss M, eds) Modern Trends in BioThermo-Kinetics 3. Innsbruck Univ Press:283-7. »Open Access

Follow-up reviews on high-resolution respirometry

• Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97. »Bioblast Access
• Hütter E, Unterluggauer H, Garedew A, Jansen-Dürr P, Gnaiger E (2006) High-resolution respirometry - a modern tool in aging research. Exp Gerontol 41:103-9. »PMID: 16309877
• Gnaiger E (2008) Polarographic oxygen sensors, the oxygraph and high-resolution respirometry to assess mitochondrial function. In: Mitochondrial Dysfunction in Drug-Induced Toxicity (Dykens JA, Will Y, eds) John Wiley:327-52. »Bioblast Access
• Pesta D, Gnaiger E (2012) High-resolution respirometry. OXPHOS protocols for human cells and permeabilized fibres from small biopisies of human muscle. Methods Mol Biol 810:25-58. »Bioblast Access

Follow-up reviews on mitochondrial and cellular oxygen kinetics

• Gnaiger E, Lassnig B, Kuznetsov AV, Margreiter R (1998) Mitochondrial respiration in the low oxygen environment of the cell: Effect of ADP on oxygen kinetics. Biochim Biophys Acta 1365:249-54. »PMID: 9693739
• Gnaiger E, Lassnig B, Kuznetsov AV, Rieger G, Margreiter R (1998) Mitochondrial oxygen affinity, respiratory flux control, and excess capacity of cytochrome c oxidase. J Exp Biol 201:1129-39. »Open Access
• Gnaiger E, Méndez G, Hand SC (2000) High phosphorylation efficiency and depression of uncoupled respiration in mitochondria under hypoxia. Proc Natl Acad Sci U S A 97:11080-5. »Open Access
• Gnaiger E (2001) Bioenergetics at low oxygen: dependence of respiration and phosphorylation on oxygen and adenosine diphosphate supply. Respir Physiol 128:277-97. »Bioblast Access
• Gnaiger E, Kuznetsov AV (2002) Mitochondrial respiration at low levels of oxygen and cytochrome c. Biochem Soc Trans 30:242-8. »PMID: 12023860
• Gnaiger E (2003) Oxygen conformance of cellular respiration. A perspective of mitochondrial physiology. Adv Exp Med Biol 543:39-55. »Bioblast Access
• Scandurra FM, Gnaiger E (2010) Cell respiration under hypoxia: facts and artefacts in mitochondrial oxygen kinetics. Adv Exp Med Biol 662:7-25. »Bioblast Access