Periodic fluctuations in oxygen consumption comparing HeLa (cancer) and CHO (non-cancer) cells and response to external NAD(P)+/NAD(P)H.
Oxygen consumption in the presence of cyanide was utilized as a measure of plasma membrane electron transport in Chinese hamster ovary (CHO) and human cervical carcinoma (HeLa) cell lines. Both intact cells and isolated plasma membranes carry cyanide-insensitive NADH(P)H oxidases at their external membrane surfaces (designated ECTO-NOX proteins). Regular oscillatory patterns of oxygen consumption with period lengths characteristic of those observed for rates of NADH oxidation by ECTO-NOX proteins were observed to provide evidence for transfer of protons and electrons to reduce oxygen to water. The oscillations plus the resistance to inhibition by cyanide identify the bulk of the oxygen consumption as due to ECTO-NOX proteins. With intact CHO cells, oxygen consumption was enhanced by but not dependent upon external NAD(P)H addition. With intact HeLa cells, oxygen consumption was inhibited by both NADH and NAD+ as was growth. The results suggest that plasma membrane electron transport from internal donors to oxygen as an external acceptor is mediated through ECTO-NOX proteins and that electron transport to molecular oxygen may be differentially affected by external pyridine nucleotides depending on cell type.
Orczyk
John
J
Department of Medicinal Chemistry, Purdue University, West Lafayette, Indiana 47907-2064, USA.
Morré
Dorothy M
DM
Morré
D James
DJ
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000201:enzymology*; D006224:Cricetinae; D004579:Electron Transport; D006367:HeLa Cells; D006801:Humans; D009097:Multienzyme Complexes / Q000378:metabolism*; D009243:NAD / Q000494:pharmacology; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009249:NADP / Q000494:pharmacology; D019255:NADPH Oxidases / Q000378:metabolism*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption
D009097:Multienzyme Complexes; D009243:NAD; D009249:NADP; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; D019255:NADPH Oxidases