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Wei Chi Jinfang Ma Lixin Zhang 《Philosophical transactions of the Royal Society of London. Series B, Biological sciences》2012,367(1608):3420-3429
Major multi-protein photosynthetic complexes, located in thylakoid membranes, are responsible for the capture of light and its conversion into chemical energy in oxygenic photosynthetic organisms. Although the structures and functions of these photosynthetic complexes have been explored, the molecular mechanisms underlying their assembly remain elusive. In this review, we summarize current knowledge of the regulatory components involved in the assembly of thylakoid membrane protein complexes in photosynthetic organisms. Many of the known regulatory factors are conserved between prokaryotes and eukaryotes, whereas others appear to be newly evolved or to have expanded predominantly in eukaryotes. Their specific features and fundamental differences in cyanobacteria, green algae and land plants are discussed. 相似文献
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Olsen JD Tucker JD Timney JA Qian P Vassilev C Hunter CN 《The Journal of biological chemistry》2008,283(45):30772-30779
The mapping of the photosynthetic membrane of Rhodobacter sphaeroides by atomic force microscopy (AFM) revealed a unique organization of arrays of dimeric reaction center-light harvesting I-PufX (RC-LH1-PufX) core complexes surrounded and interconnected by light-harvesting LH2 complexes (Bahatyrova, S., Frese, R. N., Siebert, C. A., Olsen, J. D., van der Werf, K. O., van Grondelle, R., Niederman, R. A., Bullough, P. A., Otto, C., and Hunter, C. N. (2004) Nature 430, 1058-1062). However, membrane regions consisting solely of LH2 complexes were under-represented in these images because these small, highly curved areas of membrane rendered them difficult to image even using gentle tapping mode AFM and impossible with contact mode AFM. We report AFM imaging of membranes prepared from a mutant of R. sphaeroides, DPF2G, that synthesizes only the LH2 complexes, which assembles spherical intracytoplasmic membrane vesicles of approximately 53 nm diameter in vivo. By opening these vesicles and adsorbing them onto mica to form small, < or =120 nm, largely flat sheets we have been able to visualize the organization of these LH2-only membranes for the first time. The transition from highly curved vesicle to the planar sheet is accompanied by a change in the packing of the LH2 complexes such that approximately half of the complexes are raised off the mica surface by approximately 1 nm relative to the rest. This vertical displacement produces a very regular corrugated appearance of the planar membrane sheets. Analysis of the topographs was used to measure the distances and angles between the complexes. These data are used to model the organization of LH2 complexes in the original, curved membrane. The implications of this architecture for the light harvesting function and diffusion of quinones in native membranes of R. sphaeroides are discussed. 相似文献
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The ubiquity and importance of photosynthetic organisms in nature has made the molecular mechanisms of photosynthesis a widely studied subject at both structural and functional levels. A current challenge is to understand the supramolecular assembly of the proteins involved in photosynthesis in native membranes. We have used atomic force microscopy to study the architecture of the photosynthetic apparatus and analyze the structure of single molecules in chromatophores of Phaeospirillum molischianum. Core complexes are formed by the reaction center enclosed by an elliptical light harvesting complex 1. LH2 are octameric rings, assembled either with cores or in hexagonally packed LH2 antenna domains. The symmetry mismatch caused by octameric LH2 packing in a hexagonal lattice, that could be avoided in a square lattice, suggests lipophobic effects rather than specific inter-molecular interactions drive protein organization. The core and LH2 complexes are organized to form a supramolecular assembly reminiscent to that found in Rhodospirillum photometricum, and very different from that observed in Rhodobacter sphaeroides, Rb. blasticus, and Blastochloris viridis. 相似文献
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Spectroanalysis in native gels (SING): rapid spectral analysis of pigmented thylakoid membrane complexes separated by CN–PAGE
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Ben Lucker Eliezer Schwarz Sebastian Kuhlgert Elisabeth Ostendorf David M. Kramer 《The Plant journal : for cell and molecular biology》2017,92(4):744-756
Photosynthetic organisms rapidly adjust the capture, transfer and utilization of light energy to optimize the efficiency of photosynthesis and avoid photodamage. These adjustments involve fine‐tuning of expression levels and mutual interactions among electron/proton transfer components and their associated light‐harvesting antenna. Detailed studies of these interactions and their dynamics have been hindered by the low throughput and resolution of currently available research tools, which involve laborious isolation, separation and characterization steps. To address these issues, we developed an approach that measured multiple spectroscopic properties of thylakoid preparations directly in native polyacrylamide gel electrophoresis gels, enabling unprecedented resolution of photosynthetic complexes, both in terms of the spectroscopic and functional details, as well as the ability to distinguish separate complexes and thus test their functional connections. As a demonstration, we explore the thylakoid membrane components of Chlamydomonas reinhardtii acclimated to high and low light, using a combination of room temperature absorption and 77K fluorescence emission to generate a multi‐dimensional molecular and spectroscopic map of the photosynthetic apparatus. We show that low‐light‐acclimated cells accumulate a photosystem I‐containing megacomplex that is absent in high‐light‐acclimated cells and contains distinct LhcII proteins that can be distinguished based on their spectral signatures. 相似文献
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The lateral organization of biological membranes is of great importance in many biological processes, both for the formation of specific structures such as super-complexes and for function as observed in signal transduction systems. Over the last years, AFM studies, particularly of bacterial photosynthetic membranes, have revealed that certain proteins are able to segregate into functional domains with a specific organization. Furthermore, the extended non-random nature of the organization has been suggested to be important for the energy and redox transport properties of these specialized membranes. In the work reported here, using a coarse-grained Monte Carlo approach, we have investigated the nature of interaction potentials able to drive the formation and segregation of specialized membrane domains from the rest of the membrane and furthermore how the internal organization of the segregated domains can be modulated by the interaction potentials. These simulations show that long-range interactions are necessary to allow formation of membrane domains of realistic structure. We suggest that such possibly non-specific interactions may be of great importance in the lateral organization of biological membranes in general and in photosynthetic systems in particular. Finally, we consider the possible molecular origins of such interactions and suggest a fundamental role for lipid-mediated interactions in driving the formation of specialized photosynthetic membrane domains. We call these lipid-mediated interactions a ‘lipophobic effect.’ 相似文献
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Liu LN Aartsma TJ Thomas JC Lamers GE Zhou BC Zhang YZ 《The Journal of biological chemistry》2008,283(50):34946-34953
The architecture of the entire photosynthetic membrane network determines, at the supramolecular level, the physiological roles of the photosynthetic protein complexes involved. So far, a precise picture of the native configuration of red algal thylakoids is still lacking. In this work, we investigated the supramolecular architectures of phycobilisomes (PBsomes) and native thylakoid membranes from the unicellular red alga Porphyridium cruentum using atomic force microscopy (AFM) and transmission electron microscopy. The topography of single PBsomes was characterized by AFM imaging on both isolated and membrane-combined PBsomes complexes. The native organization of thylakoid membranes presented variable arrangements of PBsomes on the membrane surface. It indicates that different light illuminations during growth allow diverse distribution of PBsomes upon the isolated photosynthetic membranes from P. cruentum, random arrangement or rather ordered arrays, to be observed. Furthermore, the distributions of PBsomes on the membrane surfaces are mostly crowded. This is the first investigation using AFM to visualize the native architecture of PBsomes and their crowding distribution on the thylakoid membrane from P. cruentum. Various distribution patterns of PBsomes under different light conditions indicate the photoadaptation of thylakoid membranes, probably promoting the energy-harvesting efficiency. These results provide important clues on the supramolecular architecture of red algal PBsomes and the diverse organizations of thylakoid membranes in vivo. 相似文献
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Time-resolved (TR) infrared (IR) spectroscopy in the nanosecond to second timescale has been extensively used, in the last 30 years, in the study of photosynthetic systems. Interesting results have also been obtained at lower time resolution (minutes or even hours). In this review, we first describe the used techniques—dispersive IR, laser diode IR, rapid-scan Fourier transform (FT)IR, step-scan FTIR—underlying the advantages and disadvantages of each of them. Then, the main TR-IR results obtained so far in the investigation of photosynthetic reactions (in reaction centers, in light-harvesting systems, but also in entire membranes or even in living organisms) are presented. Finally, after the general conclusions, the perspectives in the field of TR-IR applied to photosynthesis are described. 相似文献
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Recent topographs of the intracytoplasmic membrane (ICM) of purple bacteria obtained by atomic force microscopy (AFM) have
provided the first surface views of the native architecture of a multicomponent biological membrane at submolecular resolution,
representing an important landmark in structural biology. A variety of species-dependent, closely packed arrangements of light-harvesting
(LH) complexes was revealed: the most highly organized was found in Rhodobacter sphaeroides in which the peripheral LH2 antenna was seen either in large clusters or in fixed rows interspersed among ordered arrays
of dimeric LH1-reaction center (RC) core complexes. A more random organization was observed in other species containing both
the LH1 and LH2 complexes, as typified by Rhododspirillum photometricum with randomly packed monomeric LH1-RC core complexes intermingled with large, paracrystalline domains of LH2 antenna. Surprisingly,
no structures that could be identified as the ATP synthase or cytochrome bc
1 complexes were observed, which may reflect their localization at ICM vesicle poles or in curved membrane areas, out of view
from the flat regions imaged by AFM. This possible arrangement of energy transducing complexes has required a reassessment
of energy tranduction mechanisms which place the cytochrome bc
1 complex in close association with the RC. Instead, more plausible proposals must account for the movement of quinone redox
species over considerable membrane distances on appropriate time scales. AFM, together with atomic resolution structures are
also providing the basis for molecular modeling of the ICM that is leading to an improved picture of the supramolecular organization
of photosynthetic complexes, as well as the forces that drive their segregation into distinct domains. 相似文献
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Aurlie Crepin Zuzana Ku
erov Artemis Kosta Eric Durand Stefano Caffarri 《The Plant journal : for cell and molecular biology》2020,102(2):398-409
The biological conversion of light energy into chemical energy is performed by a flexible photosynthetic machinery located in the thylakoid membranes. Photosystems I and II (PSI and PSII) are the two complexes able to harvest light. PSI is the last complex of the electron transport chain and is composed of multiple subunits: the proteins building the catalytic core complex that are well conserved between oxygenic photosynthetic organisms, and, in green organisms, the membrane light‐harvesting complexes (Lhc) necessary to increase light absorption. In plants, four Lhca proteins (Lhca1–4) make up the antenna system of PSI, which can be further extended to optimize photosynthesis by reversible binding of LHCII, the main antenna complex of photosystem II. Here, we used biochemistry and electron microscopy in Arabidopsis to reveal a previously unknown supercomplex of PSI with LHCII that contains an additional Lhca1–a4 dimer bound on the PsaB–PsaI–PsaH side of the complex. This finding contradicts recent structural studies suggesting that the presence of an Lhca dimer at this position is an exclusive feature of algal PSI. We discuss the features of the additional Lhca dimer in the large plant PSI–LHCII supercomplex and the differences with the algal PSI. Our work provides further insights into the intricate structural plasticity of photosystems. 相似文献
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Fotiadis D 《Current opinion in biotechnology》2012,23(4):510-515
Fundamental biological processes such as cell-cell communication, signal transduction, molecular transport and energy conversion are performed by membrane proteins. These important proteins are studied best in their native environment, the lipid bilayer. The atomic force microscope (AFM) is the instrument of choice to determine the native surface structure, supramolecular organization, conformational changes and dynamics of membrane-embedded proteins under near-physiological conditions. In addition, membrane proteins are imaged at subnanometer resolution and at the single molecule level with the AFM. This review highlights the major advances and results achieved on reconstituted membrane proteins and native membranes as well as the recent developments of the AFM for imaging. 相似文献
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Photosynthetic organisms drive their metabolism by converting light energy into an electrochemical gradient with high efficiency. This conversion depends on the diffusion of quinones within the membrane. In purple photosynthetic bacteria, quinones reduced by the reaction center (RC) diffuse to the cytochrome bc(1) complex and then return once reoxidized to the RC. In Rhodospirillum photometricum the RC-containing core complexes are found in a disordered molecular environment, with fixed light-harvesting complex/core complex ratio but without a fixed architecture, whereas additional light-harvesting complexes synthesized under low-light conditions pack into large paracrystalline antenna domains. Here, we have analyzed, using time-lapse atomic force microscopy, the dynamics of the protein complexes in the different membrane domains and find that the disordered regions are dynamic whereas ordered antennae domains are static. Based on our observations we propose, and analyze using Monte Carlo simulations, a model for quinone diffusion in photosynthetic membranes. We show that the formation of large static antennae domains may represent a strategy for increasing electron transfer rates between distant complexes within the membrane and thus be important for photosynthetic efficiency. 相似文献
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Roles of the acidic lipids sulfoquinovosyl diacylglycerol and phosphatidylglycerol in photosynthesis: their specificity and evolution 总被引:3,自引:0,他引:3
Sato N 《Journal of plant research》2004,117(6):495-505
Sulfoquinovosyl diacylglycerol (SQDG) and phosphatidylglycerol (PG) are lipids with negative charges, distributed among membranes of chloroplasts of plants and their postulated progenitors, cyanobacteria, and also widely among membranes of anoxygenic photosynthetic bacteria. Thus, these acidic lipids are of great interest in terms of their roles in the function and evolution of the photosynthetic membranes. The physiological significance of these lipids in photosynthesis has been examined through characterization of mutants defective in their abilities to synthesize SQDG or PG, and through characterization of isolated thylakoid membranes or photosynthetic particles, the acidic lipid contents of which were manipulated in vitro, for example, on treatment with phospholipase to degrade PG. Responsibility of SQDG or PG has been clarified so far in terms of the structural and/or functional integrity of photosystems I and/or II in cyanobacterial, green algal, and higher plant species. Also implied were distinct levels of the responsibility in the different photosynthetic organisms. Extreme cases involved the indispensability of SQDG for photosynthesis and growth in two prokaryotic, photosynthetic organisms and the contribution of PG to construction of the photosystem-I trimer exclusively in cyanobacteria. Here, roles of these acidic lipids are discussed with a focus on their specificity and the evolution of photosynthetic membranes.Norihiro Sato is the recipient of the Botanical Society Award for Young Scientist, 2003. 相似文献
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Biological membranes compartmentalize and define physical borders of cells. They are crowded with membrane proteins that fulfill diverse crucial functions. About one-third of all genes in organisms code for, and the majority of drugs target, membrane proteins. To combine structure and function analysis of membrane proteins, we designed a two-chamber atomic force microscopy (AFM) setup that allows investigation of membranes spanned over nanowells, therefore separating two aqueous chambers. We imaged nonsupported surface layers (S layers) of Corynebacterium glutamicum at sufficient resolution to delineate a 15 A-wide protein pore. We probed the elastic and yield moduli of nonsupported membranes, giving access to the lateral interaction energy between proteins. We combined AFM and fluorescence microscopy to demonstrate the functionality of proteins in the setup by documenting proton pumping by Halobacterium salinarium purple membranes. 相似文献
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State transitions reveal the dynamics and flexibility of the photosynthetic apparatus. 总被引:29,自引:0,他引:29
F A Wollman 《The EMBO journal》2001,20(14):3623-3630
The chloroplast-based photosynthetic apparatus of plants and algae associates various redox cofactors and pigments with approximately 70 polypeptides to form five major transmembrane protein complexes. Among these are two photosystems that have distinct light absorption properties but work in series to produce reducing equivalents aimed at the fixation of atmospheric carbon. A short term chromatic adaptation known as 'State transitions' was discovered thirty years ago that allows photosynthetic organisms to adapt to changes in light quality and intensity which would otherwise compromise the efficiency of photosynthetic energy conversion. A two-decade research effort has finally unraveled the major aspects of the molecular mechanism responsible for State transitions, and their physiological significance has been revisited. This review describes how a-still elusive-regulatory kinase senses the physiological state of the photosynthetic cell and triggers an extensive supramolecular reorganization of the photosynthetic membranes. The resulting picture of the photosynthetic apparatus is that of a highly flexible energy convertor that adapts to the ever-changing intracellular demand for ATP and/or reducing power. 相似文献
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Marco Mellon Mattia Storti Antoni M. Vera-Vives David M. Kramer Alessandro Alboresi Tomas Morosinotto 《Plant physiology》2021,187(2):931
Light is the ultimate source of energy for photosynthetic organisms, but respiration is fundamental for supporting metabolism during the night or in heterotrophic tissues. In this work, we isolated Physcomitrella (Physcomitrium patens) plants with altered respiration by inactivating Complex I (CI) of the mitochondrial electron transport chain by independently targeting on two essential subunits. Inactivation of CI caused a strong growth impairment even in fully autotrophic conditions in tissues where all cells are photosynthetically active, demonstrating that respiration is essential for photosynthesis. CI mutants showed alterations in the stoichiometry of respiratory complexes while the composition of photosynthetic apparatus was substantially unaffected. CI mutants showed altered photosynthesis with high activity of both Photosystems I and II, likely the result of high chloroplast ATPase activity that led to smaller ΔpH formation across thylakoid membranes, decreasing photosynthetic control on cytochrome b6f in CI mutants. These results demonstrate that alteration of respiratory activity directly impacts photosynthesis in P. patens and that metabolic interaction between organelles is essential in their ability to use light energy for growth.Mosses lacking mitochondrial Complex I show induced activation of chloroplast ATPase, demonstrating the impact of respiration on the metabolism of plant photosynthetic cells. 相似文献
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Atomic force microscopy (AFM) has developed into a powerful tool in membrane biology. AFM features an outstanding signal-to-noise ratio that allows substructures on individual macromolecules to be visualized. Most recently, AFM topographs have shown the supramolecular assembly of the bacterial photosynthetic complexes in native membranes. Here, we have determined the translational and rotational degrees of freedom of the complexes in AFM images of multi-protein assemblies, in order to build realistic atomic models of supramolecular assemblies by docking high-resolution structures into the topographs. Membrane protein assemblies of megadalton size comprising several hundreds of polypeptide chains and pigments were built with Angstrom precision. 相似文献
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Svetozar Stoichev Sashka?B. Krumova Tonya Andreeva Jon?V. Busto Svetla Todinova Konstantin Balashev Mira Busheva Félix?M. Go?i Stefka?G. Taneva 《Biophysical journal》2015,108(4):844-853
Protonation of the lumen-exposed residues of some photosynthetic complexes in the grana membranes occurs under conditions of high light intensity and triggers a major photoprotection mechanism known as energy dependent nonphotochemical quenching. We have studied the role of protonation in the structural reorganization and thermal stability of isolated grana membranes. The macroorganization of granal membrane fragments in protonated and partly deprotonated state has been mapped by means of atomic force microscopy. The protonation of the photosynthetic complexes has been found to induce large-scale structural remodeling of grana membranes—formation of extensive domains of the major light-harvesting complex of photosystem II and clustering of trimmed photosystem II supercomplexes, thinning of the membrane, and reduction of its size. These events are accompanied by pronounced thermal destabilization of the photosynthetic complexes, as evidenced by circular dichroism spectroscopy and differential scanning calorimetry. Our data reveal a detailed nanoscopic picture of the initial steps of nonphotochemical quenching. 相似文献