Separation by blue-native PAGE and identification of the whole NAD(P)H dehydrogenase complex from barley stroma thylakoids

Quiles, Marı́a José; García, Antonio Pérez; Cuello, Juan

doi:10.1016/s0981-9428(00)00740-3

Cited by 38 publications

(24 citation statements)

References 30 publications

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“…FNR (Guedeney et al,1996;Quiles and Cuello, 1998;Funk et al, 1999;Quiles et al, 2000) and nuclear-encoded homologs of NuoE, F, and G (Rasmusson et al, 1998;Quiles et al, 2003) have been suggested to be possible candidates, but no genetic or biochemical confirmation of their involvement was obtained. An alternative strategy, using a genetic approach based on Arabidopsis mutant screening and chlorophyll fluorescence imaging, has been used in recent years to identify missing Ndh complex subunits (Hashimoto et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

“…The nature of missing subunits involved in electron input associated to the peripheral arm as well as the nature of chloroplast electron donors to the complex remain unknown. It has been proposed that ferredoxin-NADP reductase (FNR) could interact with the Ndh complex to supply electrons from NADPH, thanks to its diaphorase activity (Guedeney et al, 1996), but this possibility has been a subject of controversy (Quiles and Cuello, 1998;Sazanov et al, 1998;Funk et al, 1999;Quiles et al, 2000). Genes encoding missing subunits could have been relocated to the nucleus, as is the case for major photosynthetic complexes (PSI, photosystem II, cytochrome b 6 f, and ATPase), which are partly encoded by nuclear genes (Rochaix, 2002).…”

Section: Introductionmentioning

confidence: 99%

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New Subunits NDH-M, -N, and -O, Encoded by Nuclear Genes, Are Essential for Plastid Ndh Complex Functioning in Higher Plants

et al. 2005

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In higher plants, the Ndh complex reduces plastoquinones and is involved in cyclic electron flow around photosystem I, supplying extra-ATP for photosynthesis, particularly under environmental stress conditions. Based on plastid genome sequences, the Ndh complex would contain 11 subunits (NDH-A to -K), but homologies with bacterial complex indicate the probable existence of additional subunits. To identify missing subunits, tobacco (Nicotiana tabacum) NDH-H was His tagged at its N terminus using plastid transformation. A functional Ndh subcomplex was purified by Ni 2þ affinity chromatography and its subunit composition analyzed by mass spectrometry. Five plastid encoded subunits (NDH-A, -H, -I, -J, and -K) were identified as well as three new subunits (NDH-M, -N, and -O) homologous to cyanobacterial and higher plant proteins. Arabidopsis thaliana mutants missing one of these new subunits lack a functional Ndh complex, and NDH-M and NDH-N are not detected in a tobacco transformant lacking the Ndh complex. We discuss the involvement of these three nuclear-encoded subunits in the functional integrity of the plastidial complex.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

New Subunits NDH-M, -N, and -O, Encoded by Nuclear Genes, Are Essential for Plastid Ndh Complex Functioning in Higher Plants

et al. 2005

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“…It has been reported that FNR in higher plant chloroplasts is localized peripherally on the stromal side of thylakoid membranes through association with an intrinsic protein (Vallejos et al, 1984;Matthijs et al, 1986), PSI (Andersen et al, 1992), cytochrome (cyt) b 6 f complex (Clark et al, 1984;Zhang et al, 2001), and NAD(P)H dehydrogenase complex (Quiles et al, 2000).…”

Section: In Higher Plants Ferredoxin (Fd):nadph Oxidoreductase (Fnr)mentioning

confidence: 99%

“…It is probable that not all LFNR is associated with the cyt b 6 f complex. LFNR1 and LFNR2 may also form isoenzyme-specific or nonspecific associations with PSI and the NAD(P)H dehydrogenase complex, as described by Andersen et al (1992) and Quiles et al (2000), respectively. The presence of LFNR at the cyt b 6 f complex implies a role interconnecting the linear electron transport chain with Fd-dependent cyclic electron transport.…”

Section: Quantitation Of Lfnr Isoenzymesmentioning

confidence: 99%

Three Maize Leaf Ferredoxin:NADPH Oxidoreductases Vary in Subchloroplast Location, Expression, and Interaction with Ferredoxin

et al. 2005

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In higher plants, ferredoxin (Fd):NADPH oxidoreductase (FNR) catalyzes reduction of NADP1 in the final step of linear photosynthetic electron transport and is also implicated in cyclic electron flow. We have identified three leaf FNR isoenzymes (LFNR1, LFNR2, and LFNR3) in maize (Zea mays) chloroplasts at approximately equivalent concentrations. Fractionation of chloroplasts showed that, while LFNR3 is an exclusively soluble enzyme, LFNR1 is only found at the thylakoid membrane and LFNR2 has a dual location. LFNR1 and LFNR2 were found to associate with the cytochrome b 6 f complex following its partial purification. We cloned LFNR3 and produced all three isoenzymes as stable, soluble proteins. Measurement of Fd reduction ability showed no significant differences between these recombinant enzymes. Column chromatography revealed variation between the interaction mechanisms of LFNR1 and LFNR2 with Fd, as detected by differential dependence on specific intermolecular salt bridges and variable sensitivity of interactions to changes in pH. A comparison of LFNR transcripts in leaves of plants grown on variable nitrogen regimes revealed that LFNR1 and LFNR2 transcripts are relatively more abundant under conditions of high demand for NADPH. These results are discussed in terms of the functional differentiation of maize LFNR isoenzymes.Ferredoxin (Fd):NADPH oxidoreductase (FNR; EC 1.18.1.2) is a flavoenzyme that catalyzes reduction of NADP 1 or oxidation of NADPH through electron transfer with Fd. In the final step of photosynthetic electron transport, FNR reduces NADP 1

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“…An NADH-dehydrogenase complex (NDH) containing some ndh gene products recently has been purified from pea and barley thylakoid membranes (Sazanov et al, 1998;Quiles et al, 2000). To elucidate the function of the plastidial NDH complex in C 3 plants, ndh genes were inactivated by chloroplast transformation of tobacco (Nicotiana tabacum var Petit Havana) in different laboratories.…”

mentioning

confidence: 99%

Increased Sensitivity of Photosynthesis to Antimycin A Induced by Inactivation of the Chloroplast ndhB Gene. Evidence for a Participation of the NADH-Dehydrogenase Complex to Cyclic Electron Flow around Photosystem I

et al. 2001

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) were used to study the role of the NADH-dehydrogenase complex (NDH) during photosynthesis and particularly the involvement of this complex in cyclic electron flow around photosystem I (PSI). Photosynthetic activity was determined on leaf discs by measuring CO 2 exchange and chlorophyll fluorescence quenchings during a dark-to-light transition. In the absence of treatment, both non-photochemical and photochemical fluorescence quenchings were similar in ndhB Ϫ and wild type (WT). When leaf discs were treated with 5 m antimycin A, an inhibitor of cyclic electron flow around PSI, both quenchings were strongly affected. At steady state, maximum photosynthetic electron transport activity was inhibited by 20% in WT and by 50% in ndhB Ϫ . Under non-photorespiratory conditions (2% O 2 , 2,500 L L Ϫ1 CO 2 ), antimycin A had no effect on photosynthetic activity of WT, whereas a 30% inhibition was observed both on quantum yield of photosynthesis assayed by chlorophyll fluorescence and on CO 2 assimilation in ndhB Ϫ . The effect of antimycin A on ndhB Ϫ could not be mimicked by myxothiazol, an inhibitor of the mitochondrial cytochrome bc 1 complex, therefore showing that it is not related to an inhibition of the mitochondrial electron transport chain but rather to an inhibition of cyclic electron flow around PSI. We conclude to the existence of two different pathways of cyclic electron flow operating around PSI in higher plant chloroplasts. One of these pathways, sensitive to antimycin A, probably involves ferredoxin plastoquinone reductase, whereas the other involves the NDH complex. The absence of visible phenotype in ndhB Ϫ plants under normal conditions is explained by the complement of these two pathways in the supply of extra-ATP for photosynthesis.During oxygenic photosynthesis of C 3 plants, both photosystem II (PSII) and photosystem I (PSI) cooperate to achieve NADP ϩ reduction using water as an electron donor and generate a trans-membrane proton gradient driving ATP synthesis. Although NADP ϩ reduction is recognized to be dependent on the activity of both photosystems through electron transport reactions of the "Z" scheme (Hill and Bendall, 1960;Redding et al., 1999), it has early been reported from studies on isolated thylakoids that ATP could be produced by the sole PSI through cyclic electron transfer reactions (Arnon, 1959). The cyclic electron flow around PSI has been extensively studied in thylakoids and/or chloroplasts of C 3 plants (for review, see Fork and Herbert, 1993; Bendall and Manasse, 1995). This mechanism has been suggested to provide ATP for a variety of cellular processes, including stress adaptation (Havaux et al., 1991) and CO 2 fixation (Furbank and Horton, 1987; Herbert et al., 1990). During photosynthetic CO 2 fixation, both NADPH and ATP are used to regenerate ribulose-1,5-bisphosphate and allow functioning of the photosynthetic carbon reduction cycle (Calvin cycle). In the absence of Q cycle, when one NADPH is produced by linear electron transport reactions, four H ϩ are released in ...

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Separation by blue-native PAGE and identification of the whole NAD(P)H dehydrogenase complex from barley stroma thylakoids

Cited by 38 publications

References 30 publications

New Subunits NDH-M, -N, and -O, Encoded by Nuclear Genes, Are Essential for Plastid Ndh Complex Functioning in Higher Plants

New Subunits NDH-M, -N, and -O, Encoded by Nuclear Genes, Are Essential for Plastid Ndh Complex Functioning in Higher Plants

Three Maize Leaf Ferredoxin:NADPH Oxidoreductases Vary in Subchloroplast Location, Expression, and Interaction with Ferredoxin

Increased Sensitivity of Photosynthesis to Antimycin A Induced by Inactivation of the Chloroplast ndhB Gene. Evidence for a Participation of the NADH-Dehydrogenase Complex to Cyclic Electron Flow around Photosystem I

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