Arabidopsis FNRL protein is an NADPH-dependent chloroplast oxidoreductase resembling bacterial ferredoxin-NADP+ reductases

M Koskela, Käthe Dahlström, G Goñi, N Lehtimäki, M Nurmi, A Velazquez-Campoy, G Hanke, B Bölter, Tiina Salminen, M Medina, P Mulo

Research output: Contribution to journalArticleScientificpeer-review

5 Citations (Scopus)

Abstract

Plastidic ferredoxin-NADP+ oxidoreductases (FNRs; EC:1.18.1.2) together with bacterial type FNRs (FPRs) form the plant-type FNR family. Members of this group contain a two-domain scaffold that forms the basis of an extended superfamily of flavin adenine dinucleotide (FAD) dependent oxidoreductases. In this study, we show that the Arabidopsis thaliana At1g15140 [Ferredoxin-NADP+ oxidoreductase-like (FNRL)] is an FAD-containing NADPH dependent oxidoreductase present in the chloroplast stroma. Determination of the kinetic parameters using the DCPIP NADPH-dependent diaphorase assay revealed that the reaction catalysed by a recombinant FNRL protein followed a saturation Michaelis-Menten profile on the NADPH concentration with kcat  = 3.2 ± 0.2 s-1 , KmNADPH  = 1.6 ± 0.3 μM and kcat /KmNADPH  = 2.0 ± 0.4 μM-1  s-1 . Biochemical assays suggested that FNRL is not likely to interact with Arabidopsis ferredoxin 1, which is supported by the sequence analysis implying that the known Fd-binding residues in plastidic FNRs differ from those of FNRL. In addition, based on structural modelling FNRL has an FAD-binding N-terminal domain built from a six-stranded β-sheet and one α-helix, and a C-terminal NADP+ -binding α/β domain with a five-stranded β-sheet with a pair of α-helices on each side. The FAD-binding site is highly hydrophobic and predicted to bind FAD in a bent conformation typically seen in bacterial FPRs.
Original languageUndefined/Unknown
Pages (from-to)177–190
JournalPhysiologia Plantarum
Volume162
Issue number2
DOIs
Publication statusPublished - 2018
MoE publication typeA1 Journal article-refereed

Cite this