Arginine and lysine residues as NADH-binding sites in NADH-nitrate reductase from spinach.
Chemical modifications of spinach leaf nitrate reductase, and its 28,000 M(r) fragment with phenylglyoxal, 2,3-butanedione and pyridoxal phosphate reduce the catalytic activity of the enzyme. The kinetics of the modification indicate a rapid inactivation followed by a slower rate of inactivation. NADH-nitrate reductase, NADH-cytochrome c reductase and NADH-ferricyanide reductase activities of the nitrate reductase complex are inactivated at a faster rate when compared to the loss of FMNH2-nitrate reductase and reduced methyl viologen (MVH)-nitrate reductase activities. NADH protects the inactivation of NADH-ferricyanide reductase activity of the 28,000 M(r) fragment of nitrate reductase. These data suggest that nitrate reductase contains active sites of arginine and lysine residues that are involved in the NADH binding site of the enzyme.
Department of Agricultural Chemistry, Faculty of Horticulture, Chiba University, Japan.
Y
Y
Sato
N
N
Shiraishi
T
T
Sato
N
N
Ogura
H
H
Nakagawa
D000595:Amino Acid Sequence; D001120:Arginine / Q000378:metabolism*; D001665:Binding Sites; D003931:Diacetyl / Q000494:pharmacology; D006801:Humans; D008239:Lysine / Q000378:metabolism*; D008969:Molecular Sequence Data; D009243:NAD / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases / Q000037:antagonists & inhibitors; D050895:Nitrate Reductase (NADH); D009565:Nitrate Reductases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010658:Phenylglyoxal / Q000494:pharmacology; D010944:Plants / Q000201:enzymology; D011732:Pyridoxal Phosphate / Q000494:pharmacology; D012689:Sequence Homology, Nucleic Acid
D009243:NAD; D011732:Pyridoxal Phosphate; D001120:Arginine; D009247:NADH, NADPH Oxidoreductases; C014227:ferricyanide reductase; D009565:Nitrate Reductases; D050895:Nitrate Reductase (NADH); D003931:Diacetyl; D008239:Lysine; D010658:Phenylglyoxal