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- Title
C-terminal residues of ferredoxin-NAD(P)<sup>+</sup> reductase from <italic>Chlorobaculum tepidum</italic> are responsible for reaction dynamics in the hydride transfer and redox equilibria with NADP<sup>+</sup>/NADPH.
- Authors
Seo, Daisuke; Asano, Tomoya
- Abstract
Ferredoxin-NAD(P)+ reductase ([EC 1.18.1.2], [EC 1.18.1.3]) from <italic>Chlorobaculum tepidum</italic> (<italic>Ct</italic>FNR) is structurally homologous to the bacterial NADPH-thioredoxin reductase (TrxR), but possesses a unique C-terminal extension relative to TrxR that interacts with the isoalloxazine ring moiety of the flavin adenine dinucleotide prosthetic group. In this study, we introduce truncations to the C-terminal residues to examine their role in the reactions of <italic>Ct</italic>FNR with NADP+ and NADPH by spectroscopic and kinetic analyses. The truncation of the residues from Tyr326 to Glu360 (the whole C-terminal extension region), from Phe337 to Glu360 (omitting Phe337 on the <italic>re</italic>-face of the isoalloxazine ring) and from Ser338 to Glu360 (leaving Phe337 intact) resulted in a blue-shift of the flavin absorption bands. The truncations caused a slight increase in the dissociation constant toward NADP+ and a slight decrease in the Michaelis constant toward NADPH in steady-state assays. Pre-steady-state studies of the redox reaction with NADPH demonstrated that deletions of Tyr326-Glu360 decreased the hydride transfer rate, and the amount of reduced enzyme increased at equilibrium relative to wild-type <italic>Ct</italic>FNR. In contrast, the deletions of Phe337-Glu360 and Ser338-Glu360 resulted in only slight changes in the reaction kinetics and redox equilibrium. These results suggest that the C-terminal region of <italic>Ct</italic>FNR is responsible for the formation and stability of charge-transfer complexes, leading to changes in redox properties and reactivity toward NADP+/NADPH.
- Subjects
FERREDOXINS; OXIDATION-reduction reaction; CHEMICAL reactions; NANOPARTICLES; STOICHIOMETRY
- Publication
Photosynthesis Research, 2018, Vol 136, Issue 3, p275
- ISSN
0166-8595
- Publication type
Article
- DOI
10.1007/s11120-017-0462-z