title: Cell surface oxygen consumption by mitochondrial gene knockout cells.
abstract: Mitochondrial gene knockout (rho(0)) cells that depend on glycolysis for their energy requirements show an increased ability to reduce cell-impermeable tetrazolium dyes by electron transport across the plasma membrane. In this report, we show for the first time, that oxygen functions as a terminal electron acceptor for trans-plasma membrane electron transport (tPMET) in HL60rho(0) cells, and that this cell surface oxygen consumption is associated with oxygen-dependent cell growth in the absence of mitochondrial electron transport function. Non-mitochondrial oxygen consumption by HL60rho(0) cells was extensively inhibited by extracellular NADH and NADPH, but not by NAD(+), localizing this process at the cell surface. Mitochondrial electron transport inhibitors and the uncoupler, FCCP, did not affect oxygen consumption by HL60rho(0) cells. Inhibitors of glucose uptake and glycolysis, the ubiquinone redox cycle inhibitors, capsaicin and resiniferatoxin, the flavin centre inhibitor, diphenyleneiodonium, and the NQO1 inhibitor, dicoumarol, all inhibited oxygen consumption by HL60rho(0) cells. Similarities in inhibition profiles between non-mitochondrial oxygen consumption and reduction of the cell-impermeable tetrazolium dye, WST-1, suggest that both systems may share a common tPMET pathway. This is supported by the finding that terminal electron acceptors from both pathways compete for electrons from intracellular NADH.
authors.affiliation: Department of Radiation Therapy, Wellington School of Medicine and Health Sciences, University of Otago, PO Box 7343, Wellington, New Zealand.
authors.forename: Patries M
authors.initials: PM
authors.lastname: Herst
authors.forename: An S
authors.initials: AS
authors.lastname: Tan
authors.forename: Debbie-Jane G
authors.initials: DJ
authors.lastname: Scarlett
authors.forename: Michael V
authors.initials: MV
authors.lastname: Berridge
mesh_terms: D000332:Aerobiosis; D002211:Capsaicin / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism*; D002470:Cell Survival; D001728:Dicumarol / Q000494:pharmacology; D004224:Diterpenes / Q000494:pharmacology; D004579:Electron Transport / Q000187:drug effects; D005415:Flavins / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D009861:Onium Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction; D010101:Oxygen Consumption / Q000187:drug effects*; D013778:Tetrazolium Salts / Q000494:pharmacology; D013997:Time Factors; D014451:Ubiquinone / Q000187:drug effects; D014475:Uncoupling Agents / Q000494:pharmacology
chemical_list: C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D004224:Diterpenes; D005415:Flavins; D009861:Onium Compounds; D013778:Tetrazolium Salts; D014475:Uncoupling Agents; D009243:NAD; D014451:Ubiquinone; D009249:NADP; C007517:diphenyleneiodonium; D001728:Dicumarol; C024353:resiniferatoxin; D002211:Capsaicin