title: Interaction of NADPH-adrenodoxin reductase with NADP+ as studied by pulse radiolysis.
abstract: The reduction of flavin in NADH--adrenodoxin reductase by the hydrated electron (eaq-) was investigated by pulse radiolysis. The eaq- reduced directly the flavin of the reductase to form a blue semiquinone of the enzyme. Subsequently, the semiquinone decayed by dismutation to form the oxidized and fully reduced forms of the enzyme with a second-order rate constant of 4.4 x 10(4) M-1 s-1. In the presence of equimolar NADP+, the decay of eaq- accompanied an absorption increase at 400 nm, the spectrum of which, formed transiently, is identical to that of NADP radical (NADP.). Subsequently, the transient species decayed concomitantly with the formation of the semiquinone. The rate constant in the formation of the semiquinone was independent of the concentration of the enzyme (6.1 x 10(4) s-1 at pH 7.5). From these results, it is concluded that eaq- reacts with NADP+ bound to the enzyme to form NADP. initially, and subsequently, an electron flows from the NADP. to the flavin by an intracomplex electron transfer. A similar result was obtained in the reaction of CO2- or N-methylnicotinamide radical with the NADP(+)-adrenodoxin reductase complex. These results suggest that the nicotinamide moiety of NADP+ bound to the enzyme is accessible to the solvent and masks the flavin completely.
authors.affiliation: Institute of Scientific and Industrial Research, Osaka University, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kobayashi
authors.forename: S
authors.initials: S
authors.lastname: Miura
authors.forename: M
authors.initials: M
authors.lastname: Miki
authors.forename: Y
authors.initials: Y
authors.lastname: Ichikawa
authors.forename: S
authors.initials: S
authors.lastname: Tagawa
mesh_terms: D000818:Animals; D002417:Cattle; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism*; D066298:In Vitro Techniques; D008928:Mitochondria / Q000201:enzymology; D009249:NADP / Q000378:metabolism*; D011675:Pulse Radiolysis
chemical_list: D009249:NADP; D005287:Ferredoxin-NADP Reductase