The hormone-responsive NADH oxidase of the plant plasma membrane has properties of a NADH:protein disulfide reductase.
Plasma membranes of plant cells are characterized by a plant hormone (auxin)-responsive oxidation of NADH. The latter proceeds under argon. Also, when NADH oxidation is stimulated 50% by auxin addition, oxygen consumption is reduced by 40%. These findings are reconciled by direct assays using 5,5'-dithiobis-(2nitrobenzoic acid) (DTNB) (Ellman's reagent) that show protein disulfides to be electron acceptors for auxin-stimulated NADH oxidation. In the presence of an external reducing agent such as NADH, cysteine, or dithiothreitol, protein disulfides of the membrane are reduced with a concomitant stoichiometric increase in free thiols. In the absence of an external reducing agent, or in the presence of oxidized glutathione, DTNB-reactive thiols of the plasma membrane are decreased in the presence of auxins. Several auxin-reductant combinations were effective, but the same reductants plus chemically related and growth-inactive auxin analogs were not. A cell surface location of the affected thiols demonstrated with detergents and impermeant thiol reagents suggests that the protein may have a different physiological role than oxidation of NADH. For example, it may carry out some other role more closely related to the function of the auxin hormones in cell enlargement such as protein disulfide-thiol interchange.
Chueh
P J
PJ
Department of Foods and Nutrition, Purdue University, West Lafayette, Indiana 47907, USA.
Morré
D M
DM
Penel
C
C
DeHahn
T
T
Morré
D J
DJ
D015084:2,4-Dichlorophenoxyacetic Acid / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology*; D003545:Cysteine / Q000378:metabolism; D003624:Darkness; D004220:Disulfides / Q000378:metabolism*; D004229:Dithiothreitol / Q000378:metabolism; D018546:Hypocotyl / Q000378:metabolism; D007210:Indoleacetic Acids / Q000494:pharmacology; D009097:Multienzyme Complexes / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D009247:NADH, NADPH Oxidoreductases / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D010084:Oxidation-Reduction; D010100:Oxygen / Q000378:metabolism; D010940:Plant Proteins / Q000378:metabolism*; D013025:Glycine max / Q000528:radiation effects; D013438:Sulfhydryl Compounds / Q000032:analysis
D004220:Disulfides; D007210:Indoleacetic Acids; D009097:Multienzyme Complexes; D010940:Plant Proteins; D013438:Sulfhydryl Compounds; D009243:NAD; D015084:2,4-Dichlorophenoxyacetic Acid; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; C014703:disulfide reductase (NADH); D003545:Cysteine; D010100:Oxygen; D004229:Dithiothreitol