Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 34
Filtrar
1.
Biochim Biophys Acta Bioenerg ; 1863(8): 148909, 2022 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-35952798

RESUMEN

A very high rate for cyclic electron flow (CEF) around PSI (~180 s-1 or 210 s-1 in minimum medium or in the presence of a carbon source respectively) is measured in the presence of methyl viologen (MV) in intact cells of Chlamydomonas reinhardtii under anaerobic conditions. The observation of an efficient CEF in the presence of methyl viologen is in agreement with the previous results reports of Asada et al. in broken chloroplasts (Plant Cell Physiol. 31(4) (1990) 557-564). From the analysis of the P700 and PC absorbance changes, we propose that a confinement between 2 PC molecules, 1 PSI and 1 cytb6f corresponding to a functional supercomplex is responsible for these high rates of CEF. Supercomplex formation is also observed in the absence of methyl viologen, but with lower maximal CEF rate (about 100 s-1) suggesting that this compound facilitates the mediation of electron transfer from PSI acceptors to the stromal side of cytb6f. Further analysis of CEF in mutants of Chlamydomonas defective in state transitions shows the requirement of a kinase-driven transition to state 2 to establish this functional supercomplex configuration. However, a movement of the LHCII antennae is not involved in this process. We discuss the possible involvement of auxiliary proteins, among which is a small cytb6f-associated polypeptide, the PETO protein, which is one of the targets of the STT7 kinase.


Asunto(s)
Chlamydomonas , Carbono/metabolismo , Electrones , Paraquat , Complejo de Proteína del Fotosistema I/metabolismo
2.
Biochim Biophys Acta Bioenerg ; 1862(9): 148449, 2021 09 01.
Artículo en Inglés | MEDLINE | ID: mdl-34004195

RESUMEN

Many cyanobacteria species can use both plastocyanin and cytochrome c6 as lumenal electron carriers to shuttle electrons from the cytochrome b6f to either photosystem I or the respiratory cytochrome c oxidase. In Synechocystis sp. PCC6803 placed in darkness, about 60% of the active PSI centres are bound to a reduced electron donor which is responsible for the fast re-reduction of P700in vivo after a single charge separation. Here, we show that both cytochrome c6 and plastocyanin can bind to PSI in the dark and participate to the fast phase of P700 reduction, but the fraction of pre-bound PSI is smaller in the case of cytochrome c6 than with plastocyanin. Because of the inter-connection of respiration and photosynthesis in cyanobacteria, the inhibition of the cytochrome c oxidase results in the over-reduction of the photosynthetic electron transfer chain in the dark that translates into a lag in the kinetics of P700 oxidation at the onset of light. We show that this is true both with plastocyanin and cytochrome c6, indicating that the partitioning of electron transport between respiration and photosynthesis is regulated in the same way independently of which of the two lumenal electron carriers is present, although the mechanisms of such regulation are yet to be understood.


Asunto(s)
Citocromos c6/química , Complejo de Proteína del Fotosistema I/química , Plastocianina/química , Synechocystis/metabolismo , Clorofila/química , Cianobacterias/metabolismo , Transporte de Electrón , Complejo IV de Transporte de Electrones/química , Cinética , Oxidación-Reducción , Fotosíntesis , Tilacoides/química
3.
Biochim Biophys Acta Bioenerg ; 1862(8): 148434, 2021 08 01.
Artículo en Inglés | MEDLINE | ID: mdl-33932368

RESUMEN

The chloroplast ATP synthase (CF1Fo) contains a specific feature to the green lineage: a γ-subunit redox domain that contains a cysteine couple which interacts with the torque-transmitting ßDELSEED-loop. This thiol modulation equips CF1Fo with an important environmental fine-tuning mechanism. In vitro, disulfide formation in the γ-redox domain slows down the activity of the CF1Fo at low transmembrane electrochemical proton gradient ( [Formula: see text] ), which agrees with its proposed role as chock based on recently solved structure. The γ-dithiol formation at the onset of light is crucial to maximize photosynthetic efficiency since it lowers the [Formula: see text] activation level for ATP synthesis in vitro. Here, we validate these findings in vivo by utilizing absorption spectroscopy in Arabidopsis thaliana. To do so, we monitored the [Formula: see text] present in darkness and identified its mitochondrial sources. By following the fate and components of light-induced extra [Formula: see text] , we estimated the ATP lifetime that lasted up to tens of minutes after long illuminations. Based on the relationship between [Formula: see text] and CF1Fo activity, we conclude that the dithiol configuration in vivo facilitates photosynthesis by driving the same ATP synthesis rate at a significative lower [Formula: see text] than in the γ-disulfide state. The presented in vivo findings are an additional proof of the importance of CF1Fo thiol modulation, reconciling biochemical in vitro studies and structural insights.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , ATPasas de Translocación de Protón de Cloroplastos/metabolismo , Fotosíntesis , Hojas de la Planta/metabolismo , Fuerza Protón-Motriz , Compuestos de Sulfhidrilo/metabolismo , Arabidopsis/crecimiento & desarrollo , Oxidación-Reducción , Hojas de la Planta/crecimiento & desarrollo
4.
Proc Natl Acad Sci U S A ; 116(43): 21900-21906, 2019 10 22.
Artículo en Inglés | MEDLINE | ID: mdl-31591197

RESUMEN

In plants, algae, and some photosynthetic bacteria, the ElectroChromic Shift (ECS) of photosynthetic pigments, which senses the electric field across photosynthetic membranes, is widely used to quantify the activity of the photosynthetic chain. In cyanobacteria, ECS signals have never been used for physiological studies, although they can provide a unique tool to study the architecture and function of the respiratory and photosynthetic electron transfer chains, entangled in the thylakoid membranes. Here, we identified bona fide ECS signals, likely corresponding to carotenoid band shifts, in the model cyanobacteria Synechococcus elongatus PCC7942 and Synechocystis sp. PCC6803. These band shifts, most likely originating from pigments located in photosystem I, have highly similar spectra in the 2 species and can be best measured as the difference between the absorption changes at 500 to 505 nm and the ones at 480 to 485 nm. These signals respond linearly to the electric field and display the basic kinetic features of ECS as characterized in other organisms. We demonstrate that these probes are an ideal tool to study photosynthetic physiology in vivo, e.g., the fraction of PSI centers that are prebound by plastocyanin/cytochrome c6 in darkness (about 60% in both cyanobacteria, in our experiments), the conductivity of the thylakoid membrane (largely reflecting the activity of the ATP synthase), or the steady-state rates of the photosynthetic electron transport pathways.


Asunto(s)
Synechococcus/metabolismo , Tilacoides/metabolismo , Transporte de Electrón , Electrofisiología , Potenciales de la Membrana , Fotosíntesis , Complejo de Proteína del Fotosistema I/metabolismo , Plastocianina/metabolismo
5.
Plant Physiol ; 179(2): 630-639, 2019 02.
Artículo en Inglés | MEDLINE | ID: mdl-30498023

RESUMEN

Whereas photosynthetic function under steady-state light conditions has been well characterized, little is known about its changes that occur in response to light fluctuations. Chlororespiration, a simplified respiratory chain, is widespread across all photosynthetic lineages, but its role remains elusive. Here, we show that chlororespiration plays a crucial role in intermittent-light conditions in the green alga Chlamydomonas reinhardtii Chlororespiration, which is localized in thylakoid membranes together with the photosynthetic electron transfer chain, involves plastoquinone reduction and plastoquinol oxidation by a Plastid Terminal Oxidase (PTOX). We show that PTOX activity is critical for growth under intermittent light, with severe growth defects being observed in a mutant lacking PTOX2, the major plastoquinol oxidase. We demonstrate that the hampered growth results from a major change in the kinetics of redox relaxation of the photosynthetic electron transfer chain during the dark periods. This change, in turn, has a dramatic effect on the physiology of photosynthesis during the light periods, notably stimulating cyclic electron flow at the expense of the linear electron flow.


Asunto(s)
Chlamydomonas reinhardtii/crecimiento & desarrollo , Chlamydomonas reinhardtii/metabolismo , Oxidorreductasas/metabolismo , Proteínas de Plantas/metabolismo , Chlamydomonas reinhardtii/genética , Complejo de Citocromo b6f/metabolismo , Oscuridad , Transporte de Electrón , Luz , Mutación , Oxidación-Reducción , Oxidorreductasas/genética , Fotosíntesis/fisiología , Complejo de Proteína del Fotosistema I/metabolismo , Proteínas de Plantas/genética , Plastoquinona/análogos & derivados , Plastoquinona/metabolismo , Tilacoides/metabolismo , Regulación hacia Arriba
6.
Science ; 360(6394): 1210-1213, 2018 06 15.
Artículo en Inglés | MEDLINE | ID: mdl-29903971

RESUMEN

Photosystems I and II convert solar energy into the chemical energy that powers life. Chlorophyll a photochemistry, using red light (680 to 700 nm), is near universal and is considered to define the energy "red limit" of oxygenic photosynthesis. We present biophysical studies on the photosystems from a cyanobacterium grown in far-red light (750 nm). The few long-wavelength chlorophylls present are well resolved from each other and from the majority pigment, chlorophyll a. Charge separation in photosystem I and II uses chlorophyll f at 745 nm and chlorophyll f (or d) at 727 nm, respectively. Each photosystem has a few even longer-wavelength chlorophylls f that collect light and pass excitation energy uphill to the photochemically active pigments. These photosystems function beyond the red limit using far-red pigments in only a few key positions.


Asunto(s)
Clorofila/análogos & derivados , Cianobacterias/efectos de la radiación , Fotosíntesis/efectos de la radiación , Complejo de Proteína del Fotosistema I/efectos de la radiación , Complejo de Proteína del Fotosistema II/efectos de la radiación , Clorofila/química , Clorofila/efectos de la radiación , Clorofila A , Cianobacterias/crecimiento & desarrollo , Cianobacterias/metabolismo , Luz , Complejo de Proteína del Fotosistema I/química , Complejo de Proteína del Fotosistema II/química
7.
Photosynth Res ; 129(1): 105-7, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27106571

RESUMEN

It is now well established that the source of oxygen in photosynthesis is water. The earliest suggestion previously known to us had come from René Bernard Wurmser (1930). Here, we highlight an earlier report by Monsieur De Fourcroy (1787), who had already discussed the broad outlines of such a hypothesis in a book on Chemistry written for women. We present here a free translation of a passage from this book, with the original text in French as an Appendix.


Asunto(s)
Química/historia , Fotosíntesis , Francia , Historia del Siglo XVIII
8.
Nature ; 524(7565): 366-9, 2015 Aug 20.
Artículo en Inglés | MEDLINE | ID: mdl-26168400

RESUMEN

Diatoms are one of the most ecologically successful classes of photosynthetic marine eukaryotes in the contemporary oceans. Over the past 30 million years, they have helped to moderate Earth's climate by absorbing carbon dioxide from the atmosphere, sequestering it via the biological carbon pump and ultimately burying organic carbon in the lithosphere. The proportion of planetary primary production by diatoms in the modern oceans is roughly equivalent to that of terrestrial rainforests. In photosynthesis, the efficient conversion of carbon dioxide into organic matter requires a tight control of the ATP/NADPH ratio which, in other photosynthetic organisms, relies principally on a range of plastid-localized ATP generating processes. Here we show that diatoms regulate ATP/NADPH through extensive energetic exchanges between plastids and mitochondria. This interaction comprises the re-routing of reducing power generated in the plastid towards mitochondria and the import of mitochondrial ATP into the plastid, and is mandatory for optimized carbon fixation and growth. We propose that the process may have contributed to the ecological success of diatoms in the ocean.


Asunto(s)
Organismos Acuáticos/metabolismo , Dióxido de Carbono/metabolismo , Diatomeas/citología , Diatomeas/metabolismo , Mitocondrias/metabolismo , Fotosíntesis , Plastidios/metabolismo , Fuerza Protón-Motriz , Adenosina Trifosfato/metabolismo , Organismos Acuáticos/citología , Organismos Acuáticos/enzimología , Organismos Acuáticos/genética , Ciclo del Carbono , Diatomeas/enzimología , Diatomeas/genética , Ecosistema , Proteínas Mitocondriales/deficiencia , Proteínas Mitocondriales/metabolismo , NADP/metabolismo , Océanos y Mares , Oxidación-Reducción , Oxidorreductasas/deficiencia , Oxidorreductasas/metabolismo , Fenotipo , Proteínas de Plantas/metabolismo
9.
Photosynth Res ; 120(1-2): 237-46, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24379133

RESUMEN

Illumination of intact cells of Rhodobacter sphaeroides under anaerobic conditions has a dual effect on the redox state of the quinone pool. A large oxidation of the quinone pool is observed during the first seconds following the illumination. This oxidation is suppressed by the addition of an uncoupler in agreement with a light-induced reverse electron transfer at the level of the complex I, present both in the non-invaginated part of the membrane and in the chromatophores. At longer dark times, this illumination increases the reducing power of the cells leading to a significant reduction of the others reaction centers (RCs). From the observation that a significant proportion of RCs could be reduced by the preillumination without affecting the numbers of charge separation for the RCs, we conclude that there is no rapid thermodynamic equilibrium between the quinones present in the non-invaginated part of the membrane and those localized in the chromatophores. Under anaerobic conditions where the chromatophores quinone pool is fully reduced, we deduce, on the basis of flash-induced fluorescence kinetics, that the reduced RCs are exclusively reoxidized by the quinone generated at the Q o site of the cyt bc 1 complex. The supramolecular association between a dimeric RC-LHI complex and one cyt bc 1 complex allows the confinement of a quinone between the RC-LHI directly associated to the cyt bc1 complex.


Asunto(s)
Luz , Rhodobacter sphaeroides/metabolismo , Anaerobiosis , Oxidación-Reducción/efectos de la radiación , Rhodobacter sphaeroides/efectos de la radiación
10.
Proc Natl Acad Sci U S A ; 110(50): 20123-8, 2013 Dec 10.
Artículo en Inglés | MEDLINE | ID: mdl-24277817

RESUMEN

Diatoms, unicellular phytoplankton that account for ∼40% of marine primary productivity, often dominate coastal and open-ocean upwelling zones. Limitation of growth and productivity by iron at low light is attributed to an elevated cellular Fe requirement for the synthesis of Fe-rich photosynthetic proteins. In the dynamic coastal environment, Fe concentrations and daily surface irradiance levels can vary by two to three orders of magnitude on short spatial and temporal scales. Although genome-wide studies are beginning to provide insight into the molecular mechanisms used by diatoms to rapidly respond to such fluxes, their functional role in mediating the Fe stress response remains uncharacterized. Here, we show, using reverse genetics, that a death-specific protein (DSP; previously named for its apparent association with cell death) in the coastal diatom Thalassiosira pseudonana (TpDSP1) localizes to the plastid and enhances growth during acute Fe limitation at subsaturating light by increasing the photosynthetic efficiency of carbon fixation. Clone lines overexpressing TpDSP1 had a lower quantum requirement for growth, increased levels of photosynthetic and carbon fixation proteins, and increased cyclic electron flow around photosystem I. Cyclic electron flow is an ATP-producing pathway essential in higher plants and chlorophytes with a heretofore unappreciated role in diatoms. However, cells under replete conditions were characterized as having markedly reduced growth and photosynthetic rates at saturating light, thereby constraining the benefits afforded by overexpression. Widespread distribution of DSP-like sequences in environmental metagenomic and metatranscriptomic datasets highlights the presence and relevance of this protein in natural phytoplankton populations in diverse oceanic regimes.


Asunto(s)
Diatomeas/genética , Hierro/análisis , Luz , Fotosíntesis/fisiología , Complejo de Proteína del Fotosistema I/metabolismo , Proteínas/genética , Biofisica , Carbono/análisis , Clonación Molecular , Diatomeas/crecimiento & desarrollo , Immunoblotting , Microscopía Fluorescente , Nitrógeno/análisis , Fotosíntesis/genética , Proteínas/fisiología
11.
Plant Cell ; 25(7): 2661-78, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23839788

RESUMEN

Chloroplasts of land plants characteristically contain grana, cylindrical stacks of thylakoid membranes. A granum consists of a core of appressed membranes, two stroma-exposed end membranes, and margins, which connect pairs of grana membranes at their lumenal sides. Multiple forces contribute to grana stacking, but it is not known how the extreme curvature at margins is generated and maintained. We report the identification of the CURVATURE THYLAKOID1 (CURT1) protein family, conserved in plants and cyanobacteria. The four Arabidopsis thaliana CURT1 proteins (CURT1A, B, C, and D) oligomerize and are highly enriched at grana margins. Grana architecture is correlated with the CURT1 protein level, ranging from flat lobe-like thylakoids with considerably fewer grana margins in plants without CURT1 proteins to an increased number of membrane layers (and margins) in grana at the expense of grana diameter in overexpressors of CURT1A. The endogenous CURT1 protein in the cyanobacterium Synechocystis sp PCC6803 can be partially replaced by its Arabidopsis counterpart, indicating that the function of CURT1 proteins is evolutionary conserved. In vitro, Arabidopsis CURT1A proteins oligomerize and induce tubulation of liposomes, implying that CURT1 proteins suffice to induce membrane curvature. We therefore propose that CURT1 proteins modify thylakoid architecture by inducing membrane curvature at grana margins.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Cloroplastos/metabolismo , Membranas Intracelulares/metabolismo , Tilacoides/metabolismo , Secuencia de Aminoácidos , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/clasificación , Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Cloroplastos/ultraestructura , Immunoblotting , Membranas Intracelulares/ultraestructura , Lípidos/análisis , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Datos de Secuencia Molecular , Mutación , Fosforilación , Fotosíntesis , Filogenia , Hojas de la Planta/genética , Hojas de la Planta/metabolismo , Proteolípidos/metabolismo , Proteolípidos/ultraestructura , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Aminoácido , Tilacoides/ultraestructura
12.
Biochim Biophys Acta ; 1827(7): 834-42, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23624348

RESUMEN

Chondrus crispus is a species of red algae that grows on rocks from the middle intertidal into the subtidal zones of the North Atlantic coasts. As such, it has to cope with strongly variable abiotic conditions. Here we studied the response of the photosynthetic apparatus of this red alga to illumination. We found that, as previously described in the case of the unicellular alga Rhodella violacea (E. Delphin et al., Plant Physiol. 118 (1998) 103-113), a single multi-turnover saturating pulse of light is sufficient to induce a strong quenching of fluorescence. To elucidate the mechanisms underlying this fluorescence quenching, we combined room temperature and 77K fluorescence measurements with absorption spectroscopy to monitor the redox state of the different electron carriers in the chain. In addition, we studied the dependence of these various observables upon the excitation wavelength. This led us to identify energy spill-over from Photosystem II to Photosystem I rather than a qE-type non-photochemical quenching as the major source of fluorescence quenching that develops upon a series of 200ms pulses of saturating light results, in line with the conclusion of Ley and Butler (Biochim. Biophys. Acta 592 (1980) 349-363) from their studies of the unicellular red alga Porphyridium cruentum. In addition, we show that the onset of this spill-over is triggered by the reduction of the plastoquinone pool.


Asunto(s)
Chondrus/metabolismo , Luz , Fotoquímica , Fotosíntesis/fisiología , Complejo de Proteína del Fotosistema I/metabolismo , Complejo de Proteína del Fotosistema II/metabolismo , Chondrus/efectos de la radiación , Fluorescencia , Oxidación-Reducción , Fotosíntesis/efectos de la radiación , Complejo de Proteína del Fotosistema I/química , Complejo de Proteína del Fotosistema I/efectos de la radiación , Complejo de Proteína del Fotosistema II/química , Complejo de Proteína del Fotosistema II/efectos de la radiación , Plastoquinona/química , Plastoquinona/metabolismo
13.
Photosynth Res ; 115(1): 55-63, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23625532

RESUMEN

The Benson-Calvin cycle enzymes are activated in vivo when disulfide bonds are opened by reduction via the ferredoxin-thioredoxin system in chloroplasts. Iodoacetamide reacts irreversibly with free -SH groups of cysteine residues and inhibits the enzymes responsible for CO2 fixation. Here, we investigate the effect of iodoacetamide on electron transport, when infiltrated into spinach leaves. Using fluorescence and absorption spectroscopy, we show that (i) iodoacetamide very efficiently blocks linear electron flow upon illumination of both photosystems (decrease in the photochemical yield of photosystem II) and (ii) iodoacetamide favors cyclic electron flow upon light excitation specific to PSI. These effects account for an NPQ formation even faster in iodoacetamide under far-red illumination than in the control under saturating light. Such an increase in NPQ is dependent upon the proton gradient across the thylakoid membrane (uncoupled by nigericin addition) and PGR5 (absent in Arabidopsis pgr5 mutant). Iodoacetamide very tightly insulates the electron current at the level of the thylakoid membrane from any electron leaks toward carbon metabolism, therefore, providing choice conditions for the study of cyclic electron flow around PSI.


Asunto(s)
Arabidopsis/efectos de los fármacos , Dióxido de Carbono/metabolismo , Yodoacetamida/farmacología , Fotosíntesis/efectos de los fármacos , Spinacia oleracea/efectos de los fármacos , Arabidopsis/fisiología , Arabidopsis/efectos de la radiación , Proteínas de Arabidopsis , Cloroplastos/efectos de los fármacos , Cloroplastos/metabolismo , Transporte de Electrón/efectos de los fármacos , Luz , Iluminación , Proteínas del Complejo del Centro de Reacción Fotosintética , Complejo de Proteína del Fotosistema I , Complejo de Proteína del Fotosistema II , Hojas de la Planta , Spinacia oleracea/fisiología , Spinacia oleracea/efectos de la radiación
14.
Biochim Biophys Acta ; 1817(12): 2140-8, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-22982477

RESUMEN

The physiological role of the plastid terminal oxidase (PTOX) involved in plastoquinol oxidation in chloroplasts has been investigated in vivo in tomato leaves. Enzyme activity was assessed by non-invasive methods based on the analysis of the kinetics of chlorophyll fluorescence changes. In the dark, the maximum PTOX rate was smaller than 1 electron per second per PSII. This value was further decreased upon light acclimation, and became almost negligible upon inhibition of the photosynthetic performances by reducing the CO(2) availability. In contrast, prolonged exposure to high light resulted in an increase of the overall PTOX activity, which was paralleled by an increased protein accumulation. Under all the conditions tested the enzyme activity always remained about two orders of magnitude lower than that of electron flux through the linear photosynthetic electron pathway. Therefore, PTOX cannot have a role of a safety valve for photogenerated electrons, while it could be involved in acclimation to high light. Moreover, by playing a major role in the control of the stromal redox poise, PTOX is also capable of modulating the balance between linear and cyclic electron flow around PSI during the deactivation phase of carbon assimilation that follows a light to dark transition.


Asunto(s)
Proteínas de Cloroplastos/metabolismo , Cloroplastos/metabolismo , Oxidorreductasas/metabolismo , Fotosíntesis , Plastoquinona/metabolismo , Plantones/metabolismo , Solanum lycopersicum/metabolismo , Western Blotting , Electrones , Fluorescencia , Cinética , Luz , Oxidación-Reducción , Plastoquinona/análogos & derivados , Plastoquinona/química , Tilacoides/metabolismo
15.
Proc Natl Acad Sci U S A ; 108(32): 13317-22, 2011 Aug 09.
Artículo en Inglés | MEDLINE | ID: mdl-21784980

RESUMEN

Cyclic electron flow is increasingly recognized as being essential in plant growth, generating a pH gradient across thylakoid membrane (ΔpH) that contributes to ATP synthesis and triggers the protective process of nonphotochemical quenching (NPQ) under stress conditions. Here, we report experiments demonstrating the importance of that ΔpH in protecting plants from stress and relating to the regulation of cyclic relative to linear flow. In leaves infiltrated with low concentrations of nigericin, which dissipates the ΔpH without significantly affecting the potential gradient, thereby maintaining ATP synthesis, the extent of NPQ was markedly lower, reflecting the lower ΔpH. At the same time, the photosystem (PS) I primary donor P700 was largely reduced in the light, in contrast to control conditions where increasing light progressively oxidized P700, due to down-regulation of the cytochrome bf complex. Illumination of nigericin-infiltrated leaves resulted in photoinhibition of PSII but also, more markedly, of PSI. Plants lacking ferredoxin (Fd) NADP oxidoreductase (FNR) or the polypeptide proton gradient regulation 5 (PGR5) also show reduction of P700 in the light and increased sensitivity to PSI photoinhibition, demonstrating that the regulation of the cytochrome bf complex (cyt bf) is essential for protection of PSI from light stress. The formation of a ΔpH is concluded to be essential to that regulation, with cyclic electron flow playing a vital, previously poorly appreciated role in this protective process. Examination of cyclic electron flow in plants with a reduced content of FNR shows that these antisense plants are less able to maintain a steady rate of this pathway. This reduction is suggested to reflect a change in the distribution of FNR from cyclic to linear flow, likely reflecting the formation or disassembly of FNR-cytochrome bf complex.


Asunto(s)
Arabidopsis/fisiología , Electrones , Arabidopsis/efectos de los fármacos , Arabidopsis/efectos de la radiación , Transporte de Electrón/efectos de los fármacos , Transporte de Electrón/efectos de la radiación , Cinética , Luz , Modelos Biológicos , Nigericina/farmacología , Oxidación-Reducción/efectos de los fármacos , Oxidación-Reducción/efectos de la radiación , Fotosíntesis/efectos de los fármacos , Fotosíntesis/efectos de la radiación , Complejo de Proteína del Fotosistema II/metabolismo , Teoría Cuántica
16.
Proc Natl Acad Sci U S A ; 107(28): 12728-33, 2010 Jul 13.
Artículo en Inglés | MEDLINE | ID: mdl-20616026

RESUMEN

In plants, the major route for dissipating excess light is the nonphotochemical quenching of absorbed light (NPQ), which is associated with thylakoid lumen acidification. Our data offer an interpretation for the complex relationship between changes in luminal pH and the NPQ response. Upon steady-state illumination, fast NPQ relaxation in the dark reflects the equilibration between the electrochemical proton gradient established in the light and the cellular ATP/ADP+Pi ratio. This is followed by a slower phase, which reflects the decay of the proton motive force at equilibrium, due to gradual cellular ATP consumption. In transient conditions, a sustained lag appears in both quenching onset and relaxation, which is modulated by the size of the antenna complexes of photosystem II and by cyclic electron flow around photosystem I. We propose that this phenomenon reflects the signature of protonation of specific domains in the antenna and of slow H(+) diffusion in the different domains of the chloroplast.


Asunto(s)
Fluorescencia , Protones , Cloroplastos/genética , Cloroplastos/metabolismo , Cinética , Luz , Complejo de Proteína del Fotosistema I/genética , Complejo de Proteína del Fotosistema II/genética , Complejo de Proteína del Fotosistema II/metabolismo , Plantas/genética , Plantas/metabolismo , Fuerza Protón-Motriz/genética
17.
Biochim Biophys Acta ; 1777(7-8): 676-83, 2008.
Artículo en Inglés | MEDLINE | ID: mdl-18455502

RESUMEN

We have developed a new method to quantify the transmembrane electrochemical proton gradient present in chloroplasts of dark-adapted leaves. When a leaf is illuminated by a short pulse of intense light, we observed that the light-induced membrane potential changes, measured by the difference of absorption (520 nm-546 nm), reach a maximum value (approximately 190 mV) determined by ion leaks that occur above a threshold level of the electrochemical proton gradient. After the light-pulse, the decay of the membrane potential follows a multiphasic kinetics. A marked slowdown of the rate of membrane potential decay occurs approximately 100 ms after the light-pulse, which has been previously interpreted as reflecting the switch from an activated to an inactivated state of the ATP synthase (Junge, W., Rumberg, B. and Schröder, H., Eur. J. Biochem. 14 (1970) 575-581). This transition occurs at approximately 110 mV, thereby providing a second reference level. On this basis, we have estimated the Delta micro (H(+)) level that pre-exists in the dark. Depending upon the physiological state of the leaf, this level varies from 40 to 70 mV. In the dark, the Delta micro (H(+)) collapses upon addition of inhibitors of the respiratory chain, thus showing that it results from the hydrolysis of ATP of mitochondrial origin. Illumination of the leaf for a period longer than several seconds induces a long-lived Delta micro (H(+)) increase (up to approximately 150 mV) that reflects the light-induced increase in ATP concentration. Following the illumination, Delta micro (H(+)) relaxes to its dark-adapted value according a multiphasic kinetics that is completed in more than 1 h. In mature leaf, the deactivation of the Benson-Calvin cycle follows similar kinetics as Delta micro (H(+)) decay, showing that its state of activation is mainly controlled by ATP concentration.


Asunto(s)
ATPasas de Translocación de Protón Mitocondriales/metabolismo , Hojas de la Planta/enzimología , Aclimatación , Adenosina Trifosfato/biosíntesis , Oscuridad , Electroquímica , Activación Enzimática , Luz , Potenciales de la Membrana , ATPasas de Translocación de Protón Mitocondriales/química , Hojas de la Planta/fisiología , Hojas de la Planta/efectos de la radiación , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , ATPasas de Translocación de Protón/química , ATPasas de Translocación de Protón/metabolismo
18.
Cell ; 132(2): 273-85, 2008 Jan 25.
Artículo en Inglés | MEDLINE | ID: mdl-18243102

RESUMEN

During photosynthesis, two photoreaction centers located in the thylakoid membranes of the chloroplast, photosystems I and II (PSI and PSII), use light energy to mobilize electrons to generate ATP and NADPH. Different modes of electron flow exist, of which the linear electron flow is driven by PSI and PSII, generating ATP and NADPH, whereas the cyclic electron flow (CEF) only generates ATP and is driven by the PSI alone. Different environmental and metabolic conditions require the adjustment of ATP/NADPH ratios and a switch of electron distribution between the two photosystems. With the exception of PGR5, other components facilitating CEF are unknown. Here, we report the identification of PGRL1, a transmembrane protein present in thylakoids of Arabidopsis thaliana. Plants lacking PGRL1 show perturbation of CEF, similar to PGR5-deficient plants. We find that PGRL1 and PGR5 interact physically and associate with PSI. We therefore propose that the PGRL1-PGR5 complex facilitates CEF in eukaryotes.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Complejo de Proteína del Fotosistema I/metabolismo , Tilacoides/química , Adenosina Trifosfato/biosíntesis , Secuencia de Aminoácidos , Arabidopsis/genética , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Cloroplastos/metabolismo , ADN de Plantas/genética , ADN de Plantas/aislamiento & purificación , Transporte de Electrón , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Cinética , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Modelos Biológicos , Datos de Secuencia Molecular , Mutación , NADP/biosíntesis , Oxidación-Reducción , Proteínas del Complejo del Centro de Reacción Fotosintética/genética , Plastoquinona/metabolismo , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estructura Secundaria de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/metabolismo , Homología de Secuencia de Aminoácido , Fracciones Subcelulares/metabolismo
19.
Biochim Biophys Acta ; 1767(10): 1252-9, 2007 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-17803955

RESUMEN

The pgr5 mutant of Arabidopsis thaliana has been described as being deficient in cyclic electron flow around photosystem I, however, the precise role of the PGR5 protein remains unknown. To address this issue, photosynthetic electron transport was examined in intact leaves of pgr5 and wild type A. thaliana. Based on measurements of the kinetics of P700 oxidation in far red light and re-reduction following oxidation in the presence of DCMU, we conclude that this mutant is able to perform cyclic electron flow at a rate similar to the wild type. The PGR5 protein is therefore not essential for cyclic flow. However, cyclic flow is affected by the pgr5 mutation under conditions where this process is normally enhanced in wild type leaves, i.e. high light or low CO(2) concentrations resulted in enhancement of cyclic electron flow. This suggests a different capacity to regulate cyclic flow in response to environmental stimuli in the mutant. We also show that the pgr5 mutant is affected in the redox poising of the chloroplast, with the electron transport chain being substantially reduced under most conditions. This may result in defective feedback regulation of photosynthetic electron transport under some conditions, thus providing a rationale for the reduced efficiency of cyclic electron flow.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Fotosíntesis , Proteínas del Complejo del Centro de Reacción Fotosintética/metabolismo , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Dióxido de Carbono/farmacología , Clorofila/metabolismo , Diurona/farmacología , Transporte de Electrón , Regulación de la Expresión Génica de las Plantas , Cinética , Luz , Mutación/genética , Oxidación-Reducción , Proteínas del Complejo del Centro de Reacción Fotosintética/genética , Plantas Modificadas Genéticamente , Sensibilidad y Especificidad , Células del Estroma/metabolismo
20.
Biochim Biophys Acta ; 1767(1): 56-65, 2007 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-17123461

RESUMEN

In photosynthetic chains, the kinetics of fluorescence yield depends on the photochemical rates at the level of both Photosystem I and II and thus on the absorption cross section of the photosynthetic units as well as on the coupling between light harvesting complexes and photosynthetic traps. A new set-up is described which, at variance with the commonly used set-ups, uses of a weakly absorbed light source (light-emitting diodes with maximum output at 520 nm) to excite the photosynthetic electron chain and probe the resulting fluorescence yield changes and their time course. This approach optimizes the homogeneity of the exciting light throughout the leaf and we show that this homogeneity narrows the distribution of the photochemical rates. Although the exciting light is weakly absorbed, the possibility to tune the intensity of the light emitting diodes allows one to reach photochemical rates ranging from 10(4) s(-1) to 0.25 s(-1) rendering experimentally accessible different functional regimes. The variations of the fluorescence yield induced by the photosynthetic activity are qualitatively and quantitatively discussed. When illuminating dark-adapted leaves by a weak light, the kinetics of fluorescence changes displays a pronounced plateau which precedes the fluorescence increase reflecting the full reduction of the plastoquinone pool. We ascribe this plateau to the time delay needed to reduce the photosystem I electron acceptors.


Asunto(s)
Color , Fluorescencia , Luz , Fotoquímica/instrumentación , Fotoquímica/métodos , Fotosíntesis/fisiología , Complejo de Proteína del Fotosistema I/metabolismo , Hojas de la Planta/fisiología , Cinética , Hojas de la Planta/metabolismo , Plastoquinona/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA