Growth of rho 0 human Namalwa cells lacking oxidative phosphorylation can be sustained by redox compounds potassium ferricyanide or coenzyme Q10 putatively acting through the plasma membrane oxidase.
Pyruvate is conventionally used as a key growth supplement for mammalian rho 0 cells that lack mitochondrial DNA and are thereby devoid of oxidative phosphorylation. We have tested the proposition that cultured rho 0 human cells can be grown using redox compounds other than pyruvate. The results show that potassium ferricyanide and coenzyme Q10 can each be used to replace pyruvate to support the growth of rho 0 Namalwa cells (a lymphoblastoid cell line). Ferricyanide and coenzyme Q10 have both been reported as substrates for a plasma membrane NADH oxidase system which is capable of re-oxidising cytosolic NADH to NAD+. These compounds are also known to stimulate the activity of this enzyme system. We interpret our data to indicate that redox support for growth of rho 0 human cells can be achieved by external electron acceptors such as ferricyanide (a plasma membrane impermeant compound), or coenzyme Q10 (an integral component of the plasma membrane oxidase), through the enhanced conversion of cytosolic NADH to NAD+. This re-oxidation of NADH enables glycolysis to function efficiently as the sole source of cellular ATP, in the absence of mitochondrial oxidative phosphorylation in rho 0 cells. This has important implications for the development of new strategies for the amelioration of the bioenergy decline that occurs in mitochondrial disease and during the human ageing process.
Martinus
R D
RD
Department of Biochemistry, Monash University, Clayton, Victoria, Australia.
Linnane
A W
AW
Nagley
P
P
D002455:Cell Division / Q000187:drug effects* / Q000502:physiology; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology*; D003067:Coenzymes; D003470:Culture Media; D004272:DNA, Mitochondrial / Q000502:physiology; D005292:Ferricyanides / Q000494:pharmacology*; D006801:Humans; D009097:Multienzyme Complexes / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D010085:Oxidative Phosphorylation; D011773:Pyruvates / Q000494:pharmacology; D019289:Pyruvic Acid; D014451:Ubiquinone / Q000031:analogs & derivatives* / Q000494:pharmacology
D003067:Coenzymes; D003470:Culture Media; D004272:DNA, Mitochondrial; D005292:Ferricyanides; D009097:Multienzyme Complexes; D011773:Pyruvates; D014451:Ubiquinone; D019289:Pyruvic Acid; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; C024989:coenzyme Q10; C028033:potassium ferricyanide