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, pO2, is low, typically one to two orders of magnitude below incubation conditions used routinely for the study of respiratory control in isolated mitochondria. The pO2 range of respiratory control by oxygen overlaps with cellular oxygen profiles, indicating the significance of pO2 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 pO2. Oxygen pressure for half-maximum respiration, p50, in isolated mitochondria at state 4 was 0.025 kPa (0.2 Torr; 0.3 microM 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/pO2 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.