O(2)-sensing Signal Cascade: Clamping of O(2) Respiration, Reduced ATP Utilization, and Inducible Fumarate Respiration

Overview

Journal Am J Physiol Cell Physiol

Publisher American Physiological Society

Specialties Cell Biology
Physiology

Date 2008 May 9

PMID 18463229

Citations 18

Authors

Vijayalakshmi Sridharan

Jason Guichard

Chuan-Yuan Li

Robin Muise-Helmericks

Craig Cano Beeson

Gary L Wright

Affiliations

Soon will be listed here.

Abstract

These studies explore the consequences of activating the prolyl hydroxylase (PHD) O(2)-sensing pathway in spontaneously twitching neonatal cardiomyocytes. Full activation of the PHD pathway was achieved using the broad-spectrum PHD inhibitor (PHI) dimethyloxaloylglycine (DMOG). PHI treatment of cardiomyocytes caused an 85% decrease in O(2) consumption and a 300% increase in lactic acid production under basal conditions. This indicates a approximately 75% decrease in ATP turnover rate, inasmuch as the increased ATP generation by glycolysis is inadequate to compensate for the lower respiration. To determine the extent to which decreased ATP turnover underlies the suppressed O(2) consumption, mitochondria were uncoupled with 2,4-dinitrophenol. We were surprised to find that 2,4-dinitrophenol failed to increase O(2) consumption by PHI-treated cells, indicating that electron transport chain activity, rather than ATP turnover rate, limits respiration in PHI-treated cardiomyocytes. Silencing of hypoxia-inducible factor-1alpha (HIF-1alpha) expression restored the ability of uncoupled PHI-treated myocytes to increase O(2) consumption; however, basal O(2) uptake rates remained low because of the unabated suppression of cellular ATP consumption. Thus it appears that respiration is actively "clamped" through an HIF-dependent mechanism, whereas HIF-independent mechanisms are responsible for downregulation of ATP consumption. In addition, we find that PHD pathway activation enables mitochondria to utilize fumarate as a terminal electron acceptor when cytochrome c oxidase is inactive. The source of fumarate for this unusual respiration is derived from aspartate via the purine nucleotide cycle. In sum, these studies show that the O(2)-sensing pathway is sufficient to actively "clamp" O(2) consumption and independently suppress cellular ATP consumption. The PHD pathway also enables the mitochondria to utilize fumarate for respiration.

Citing Articles

H NMR metabonomic analysis of serum and flap tissue on the effect of ginsenoside Rb1 on survival of random pattern skin flaps in rats.

Huang Y, Liu Y, Meng F, Wijaya W, Cao C Sci Rep. 2025; 15(1):7416.

PMID: 40033034 PMC: 11876603. DOI: 10.1038/s41598-025-91798-z.

High ambient temperature exposure during late gestation disrupts glycolipid metabolism and hepatic mitochondrial function tightly related to gut microbial dysbiosis in pregnant mice.

He J, Liu R, Zheng W, Guo H, Yang Y, Zhao R Microb Biotechnol. 2021; 14(5):2116-2129.

PMID: 34272826 PMC: 8449678. DOI: 10.1111/1751-7915.13893.

ALKBH7 mediates necrosis via rewiring of glyoxal metabolism.

Kulkarni C, Nadtochiy S, Kennedy L, Zhang J, Chhim S, Alwaseem H Elife. 2020; 9.

PMID: 32795389 PMC: 7442491. DOI: 10.7554/eLife.58573.

Succinate accumulation induces mitochondrial reactive oxygen species generation and promotes status epilepticus in the kainic acid rat model.

Zhang Y, Zhang M, Zhu W, Yu J, Wang Q, Zhang J Redox Biol. 2019; 28:101365.

PMID: 31707354 PMC: 6854095. DOI: 10.1016/j.redox.2019.101365.

Modelling pyruvate dehydrogenase under hypoxia and its role in cancer metabolism.

Eyassu F, Angione C R Soc Open Sci. 2017; 4(10):170360.

PMID: 29134060 PMC: 5666243. DOI: 10.1098/rsos.170360.

References

Hohl C, Oestreich R, Rosen P, Wiesner R, Grieshaber M . Evidence for succinate production by reduction of fumarate during hypoxia in isolated adult rat heart cells. Arch Biochem Biophys. 1987; 259(2):527-35. DOI: 10.1016/0003-9861(87)90519-4. View

Di Lisa F, Bernardi P . Mitochondrial function as a determinant of recovery or death in cell response to injury. Mol Cell Biochem. 1998; 184(1-2):379-91. View

Taegtmeyer H, Peterson M, Ragavan V, Ferguson A, Lesch M . De novo alanine synthesis in isolated oxygen-deprived rabbit myocardium. J Biol Chem. 1977; 252(14):5010-8. View

Heusch G, Schulz R . Myocardial hibernation. Ital Heart J. 2002; 3(5):282-4. View

Hochachka P, Storey K . Metabolic consequences of diving in animals and man. Science. 1975; 187(4177):613-21. DOI: 10.1126/science.163485. View

Epstein A, Gleadle J, McNeill L, Hewitson K, ORourke J, Mole D . C. elegans EGL-9 and mammalian homologs define a family of dioxygenases that regulate HIF by prolyl hydroxylation. Cell. 2001; 107(1):43-54. DOI: 10.1016/s0092-8674(01)00507-4. View

Papandreou I, Cairns R, Fontana L, Lim A, Denko N . HIF-1 mediates adaptation to hypoxia by actively downregulating mitochondrial oxygen consumption. Cell Metab. 2006; 3(3):187-97. DOI: 10.1016/j.cmet.2006.01.012. View

Wiesner R, Rosen P, Grieshaber M . Pathways of succinate formation and their contribution to improvement of cardiac function in the hypoxic rat heart. Biochem Med Metab Biol. 1988; 40(1):19-34. DOI: 10.1016/0885-4505(88)90100-4. View

Sridharan V, Guichard J, Bailey R, Kasiganesan H, Beeson C, Wright G . The prolyl hydroxylase oxygen-sensing pathway is cytoprotective and allows maintenance of mitochondrial membrane potential during metabolic inhibition. Am J Physiol Cell Physiol. 2006; 292(2):C719-28. DOI: 10.1152/ajpcell.00100.2006. View

10.

Sanborn T, Gavin W, Berkowitz S, Perille T, Lesch M . Augmented conversion of aspartate and glutamate to succinate during anoxia in rabbit heart. Am J Physiol. 1979; 237(5):H535-41. DOI: 10.1152/ajpheart.1979.237.5.H535. View

11.

LAPLANTE A, Vincent G, Poirier M, Des Rosiers C . Effects and metabolism of fumarate in the perfused rat heart. A 13C mass isotopomer study. Am J Physiol. 1997; 272(1 Pt 1):E74-82. DOI: 10.1152/ajpendo.1997.272.1.E74. View

12.

Elvidge G, Glenny L, Appelhoff R, Ratcliffe P, Ragoussis J, Gleadle J . Concordant regulation of gene expression by hypoxia and 2-oxoglutarate-dependent dioxygenase inhibition: the role of HIF-1alpha, HIF-2alpha, and other pathways. J Biol Chem. 2006; 281(22):15215-26. DOI: 10.1074/jbc.M511408200. View

13.

Kasiganesan H, Sridharan V, Wright G . Prolyl hydroxylase inhibitor treatment confers whole-animal hypoxia tolerance. Acta Physiol (Oxf). 2007; 190(2):163-9. DOI: 10.1111/j.1748-1716.2007.01676.x. View

14.

Semenza G . Expression of hypoxia-inducible factor 1: mechanisms and consequences. Biochem Pharmacol. 1999; 59(1):47-53. DOI: 10.1016/s0006-2952(99)00292-0. View

15.

Hochachka P, Owen T, Allen J, Whittow G . Multiple end products of anaerobiosis in diving vertebrates. Comp Biochem Physiol B. 1975; 50(1):17-22. DOI: 10.1016/0305-0491(75)90292-8. View

16.

Arthur P, Giles J, Wakeford C . Protein synthesis during oxygen conformance and severe hypoxia in the mouse muscle cell line C2C12. Biochim Biophys Acta. 2000; 1475(1):83-9. DOI: 10.1016/s0304-4165(00)00046-5. View

17.

Reers M, Smiley S, Chen A, Lin M, Chen L . Mitochondrial membrane potential monitored by JC-1 dye. Methods Enzymol. 1995; 260:406-17. DOI: 10.1016/0076-6879(95)60154-6. View

18.

Nicholls D . The influence of respiration and ATP hydrolysis on the proton-electrochemical gradient across the inner membrane of rat-liver mitochondria as determined by ion distribution. Eur J Biochem. 1974; 50(1):305-15. DOI: 10.1111/j.1432-1033.1974.tb03899.x. View

19.

Feldkamp T, Kribben A, Roeser N, Senter R, Kemner S, Venkatachalam M . Preservation of complex I function during hypoxia-reoxygenation-induced mitochondrial injury in proximal tubules. Am J Physiol Renal Physiol. 2003; 286(4):F749-59. DOI: 10.1152/ajprenal.00276.2003. View

20.

Owen T, Hochachka P . Purification and properties of dolphin muscle aspartate and alanine transaminases and thier possible roles in the energy metabolism of diving mammals. Biochem J. 1974; 143(3):541-53. PMC: 1168423. DOI: 10.1042/bj1430541. View