title: Characterization of Nda2, a plastoquinone-reducing type II NAD(P)H dehydrogenase in chlamydomonas chloroplasts.
abstract: Electron transfer pathways associated to oxygenic photosynthesis, including cyclic electron flow around photosystem I and chlororespiration, rely on non-photochemical reduction of plastoquinones (PQs). In higher plant chloroplasts, a bacterial-like NDH complex homologous to complex I is involved in PQ reduction, but such a complex is absent from Chlamydomonas plastids where a type II NAD(P)H dehydrogenase activity has been proposed to operate. With the aim to elucidate the nature of the enzyme-supporting non-photochemical reduction of PQs, one of the type II NAD(P)H dehydrogenases identified in the Chlamydomonas reinhardtii genome (Nda2) was produced as a recombinant protein in Escherichia coli and further characterized. As many type II NAD(P)H dehydrogenases, Nda2 uses NADH as a preferential substrate, but in contrast to the eukaryotic enzymes described so far, contains non-covalently bound FMN as a cofactor. When expressed at a low level, Nda2 complements growth of an E. coli lacking both NDH-1 and NDH-2, but is toxic at high expression levels. Using an antibody raised against the recombinant protein and based on its mass spectrometric identification, we show that Nda2 is localized in thylakoid membranes. Chlorophyll fluorescence measurements performed on thylakoid membranes show that Nda2 is able to interact with thylakoid membranes of C. reinhardtii by reducing PQs from exogenous NADH or NADPH. We discuss the possible involvement of Nda2 in cyclic electron flow around PSI, chlororespiration, and hydrogen production.
authors.affiliation: CEA, CEA Cadarache, Direction des Sciences du Vivant, Institut de Biologie Environnementale et de Biotechnologie, CNRS, UMR Biologie Végétale et Microbiologie Environnementale, Aix Marseille Université, F-13108 Saint-Paul-lez-Durance, France.
authors.forename: Carine
authors.initials: C
authors.lastname: Desplats
authors.forename: Florence
authors.initials: F
authors.lastname: Mus
authors.forename: Stéphan
authors.initials: S
authors.lastname: Cuiné
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Billon
authors.forename: Laurent
authors.initials: L
authors.lastname: Cournac
authors.forename: Gilles
authors.initials: G
authors.lastname: Peltier
mesh_terms: D000818:Animals; D016825:Chlamydomonas reinhardtii / Q000201:enzymology* / Q000235:genetics; D004579:Electron Transport / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics; D005816:Genetic Complementation Test; D018503:Genome, Protozoan / Q000502:physiology*; D006859:Hydrogen / Q000378:metabolism; D009249:NADP / Q000235:genetics / Q000378:metabolism; D009252:NADPH Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D010084:Oxidation-Reduction; D010971:Plastoquinone / Q000378:metabolism*; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020524:Thylakoids / Q000235:genetics / Q000378:metabolism*
chemical_list: D015800:Protozoan Proteins; D011994:Recombinant Proteins; D009249:NADP; D006859:Hydrogen; D009252:NADPH Dehydrogenase; D010971:Plastoquinone