Purification and characterization of an NAD(P)H:quinone oxidoreductase from Glycine max seedlings.
An NAD(P)H:(quinone acceptor) oxidoreductase (EC 1.6.99.2) was purified from Glycine max seedlings by means of chromatographic procedures. After 1371-fold purification, the enzyme showed a single band in IEF corresponding to an isoelectric point of 6.1. A single band was also found in native-PAGE both by activity staining and Coomassie brilliant blue staining. The molecular mass determined in SDS-PAGE was 21900 Da, while in HPLC gel-filtration it was 61000 Da. The NAD(P)H:quinone oxidoreductase was able to use NADH or NADPH as the electron donor. Among the artificial quinones which are reduced by this enzyme, 6-hydroxydopa- and 6-hydroxydopamine-quinone are of particular interest because of their neurotoxic effects.
Rescigno
A
A
Istituto di Chimica Biologica, Università di Cagliari, Italy.
Sollai
F
F
Masala
S
S
Porcu
M C
MC
Sanjust
E
E
Rinaldi
A C
AC
Curreli
N
N
Grifi
D
D
Rinaldi
A
A
D015086:2,6-Dichloroindophenol / Q000378:metabolism; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D010944:Plants / Q000201:enzymology; D013025:Glycine max / Q000201:enzymology*; D013053:Spectrophotometry; D013379:Substrate Specificity; D013696:Temperature
D009243:NAD; D009249:NADP; D015086:2,6-Dichloroindophenol; D016660:NAD(P)H Dehydrogenase (Quinone)