A growth factor- and hormone-stimulated NADH oxidase from rat liver plasma membrane.
NADH oxidase activity (electron transfer from NADH to molecular oxygen) of plasma membranes purified from rat liver was characterized by a cyanide-insensitive rate of 1 to 5 nmol/min per mg protein. The activity was stimulated by growth factors (diferric transferrin and epidermal growth factor) and hormones (insulin and pituitary extract) 2- to 3-fold. In contrast, NADH oxidase was inhibited up to 80% by several agents known to inhibit growth or induce differentiation (retinoic acid, calcitriol, and the monosialoganglioside, GM3). The growth factor-responsive NADH oxidase of isolated plasma membranes was not inhibited by common inhibitors of oxidoreductases of endoplasmic reticulum or mitochondria. As well, NADH oxidase of the plasma membrane was stimulated by concentrations of detergents which strongly inhibited mitochondrial NADH oxidases and by lysolipids or fatty acids. Growth factor-responsive NADH oxidase, however, was inhibited greater than 90% by chloroquine and quinone analogues. Addition of coenzyme Q10 stimulated the activity and partially reversed the analogue inhibition. The pH optimum for NADH oxidase was 7.0 both in the absence and presence of growth factors. The Km for NADH was 5 microM and was increased in the presence of growth factors. The stoichiometry of the electron transfer reaction from NADH to oxygen was 2 to 1, indicating a 2 electron transfer. NADH oxidase was separated from NADH-ferricyanide reductase, also present at the plasma membrane, by ion exchange chromatography. Taken together, the evidence suggests that NADH oxidase of the plasma membrane is a unique oxidoreductase and may be important to the regulation of cell growth.
Brightman
A O
AO
Department of Medicinal Chemistry, Purdue University, West Lafayette, IN 47907.
Wang
J
J
Miu
R K
RK
Sun
I L
IL
Barr
R
R
Crane
F L
FL
Morré
D J
DJ
D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology / Q000648:ultrastructure; D003902:Detergents; D004789:Enzyme Activation; D004815:Epidermal Growth Factor / Q000494:pharmacology*; D005732:Gangliosides / Q000494:pharmacology; D006728:Hormones / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D019787:Linoleic Acid; D008041:Linoleic Acids / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008854:Microscopy, Electron; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010084:Oxidation-Reduction; D051381:Rats; D014168:Transferrin / Q000494:pharmacology*; D014451:Ubiquinone / Q000494:pharmacology
D003902:Detergents; D005732:Gangliosides; D006728:Hormones; D008041:Linoleic Acids; D008244:Lysophosphatidylcholines; D009097:Multienzyme Complexes; D014168:Transferrin; C028283:diferric transferrin; D014451:Ubiquinone; D004815:Epidermal Growth Factor; D019787:Linoleic Acid; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases