Rat liver NAD(P)H:quinone oxidoreductase: cDNA expression and site-directed mutagenesis.
Rat liver NAD(P)H:quinone oxidoreductase cDNA was cloned and expressed in a eukaryotic cell expression plasmid containing a cytomegalovirus (CMV) promoter. Transient expression of enzyme activity and RNA transcription were measured in COS7 cells. The expressed quinone reductase has kinetic properties similar to the rat liver enzyme and is inhibited by dicourmarol, a known inhibitor of NAD(P)H:quinone oxidoreductase. Site-directed mutagenesis experiments carried out using this expression system revealed possible regions involved in NAD(P)H binding.
Forrest
G L
GL
Department of Biology, Beckman Research Institute/City of Hope, Duarte, CA 91010.
Qian
J
J
Ma
J X
JX
Kaplan
W D
WD
Akman
S
S
Doroshow
J
J
Chen
S A
SA
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis; D015870:Gene Expression; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D016660:NAD(P)H Dehydrogenase (Quinone); D009536:Niacinamide / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011808:Quinone Reductases / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism
D011994:Recombinant Proteins; D009536:Niacinamide; D004247:DNA; D016660:NAD(P)H Dehydrogenase (Quinone); D011808:Quinone Reductases