Photosynthetic eukaryotes whose cells harbor plastids originating from secondary endosymbiosis of a red alga include species of major ecological and economic importance. Since utilization of solar energy relies on the efficient light-harvesting, one of the critical factors for the success of the red lineage in a range of environments is to be found in the adaptability of the light-harvesting machinery, formed by the proteins of the light-harvesting complex (LHC) family. A number of species are known to employ mainly a unique class of LHC containing red-shifted chlorophyll a (Chl a) forms absorbing above 690 nm. This appears to be an adaptation to shaded habitats. Here we present a detailed investigation of excitation energy flow in the red-shifted light-harvesting antenna of eustigmatophyte Trachydiscus minutus using time-resolved fluorescence and ultrafast transient absorption measurements. The main carotenoid in the complex is violaxanthin, hence this LHC is labeled the red-violaxanthin-Chl a protein, rVCP. Both the carotenoid-to-Chl a energy transfer and excitation dynamics within the Chl a manifold were studied and compared to the related antenna complex, VCP, that lacks the red-Chl a. Two spectrally defined carotenoid pools were identified in the red antenna, contributing to energy transfer to Chl a, mostly via S2 and hot S1 states. Also, Chl a triplet quenching by carotenoids is documented. Two separate pools of red-shifted Chl a were resolved, one is likely formed by excitonically coupled Chl a molecules. The structural implications of these observations are discussed.
- MeSH
- chlorofyl a * MeSH
- Chlorophyta fyziologie MeSH
- fluorescenční spektrometrie metody MeSH
- Heterokontophyta fyziologie MeSH
- plastidy MeSH
- přenos energie fyziologie MeSH
- Rhodophyta fyziologie MeSH
- světlosběrné proteinové komplexy chemie MeSH
- xanthofyly MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Photosystem I (PSI) is a multi-subunit integral pigment-protein complex that performs light-driven electron transfer from plastocyanin to ferredoxin in the thylakoid membrane of oxygenic photoautotrophs. In order to achieve the optimal photosynthetic performance under ambient irradiance, the absorption cross section of PSI is extended by means of peripheral antenna complexes. In eukaryotes, this role is played mostly by the pigment-protein complexes of the LHC family. The structure of the PSI-antenna supercomplexes has been relatively well understood in organisms harboring the primary plastid: red algae, green algae and plants. The secondary endosymbiotic algae, despite their major ecological importance, have so far received less attention. Here we report a detailed structural analysis of the antenna-PSI association in the stramenopile alga Nannochloropsis oceanica (Eustigmatophyceae). Several types of PSI-antenna assemblies are identified allowing for identification of antenna docking sites on the PSI core. Instances of departure of the stramenopile system from the red algal model of PSI-Lhcr structure are recorded, and evolutionary implications of these observations are discussed.
Eustigmatophyte algae represent an interesting model system for the study of the regulation of the excitation energy flow due to their use of violaxanthin both as a major light-harvesting pigment and as the basis of xanthophyll cycle. Fluorescence induction kinetics was studied in an oleaginous marine alga Nannochloropsis oceanica. Nonphotochemical fluorescence quenching was analyzed in detail with respect to the state of the cellular xanthophyll pool. Two components of nonphotochemical fluorescence quenching (NPQ), both dependent on the presence of zeaxanthin, were clearly resolved, denoted as slow and fast NPQ based on kinetics of their formation. The slow component was shown to be in direct proportion to the amount of zeaxanthin, while the fast NPQ component was transiently induced in the presence of membrane potential on subsecond timescales. The applicability of these observations to other eustigmatophyte species is demonstrated by measurements of other representatives of this algal group, both marine and freshwater.
- MeSH
- fluorescence MeSH
- fotosyntéza MeSH
- mořské řasy chemie MeSH
- Publikační typ
- časopisecké články MeSH
Chlorobaculum tepidum is a representative of green sulfur bacteria, a group of anoxygenic photoautotrophs that employ chlorosomes as the main light-harvesting structures. Chlorosomes are coupled to a ferredoxin-reducing reaction center by means of the Fenna-Matthews-Olson (FMO) protein. While the biochemical properties and physical functioning of all the individual components of this photosynthetic machinery are quite well understood, the native architecture of the photosynthetic supercomplexes is not. Here we report observations of membrane-bound FMO and the analysis of the respective FMO-reaction center complex. We propose the existence of a supercomplex formed by two reaction centers and four FMO trimers based on the single-particle analysis of the complexes attached to native membrane. Moreover, the structure of the photosynthetic unit comprising the chlorosome with the associated pool of RC-FMO supercomplexes is proposed.
- MeSH
- bakteriální proteiny chemie metabolismus ultrastruktura MeSH
- Chlorobi chemie MeSH
- cytoplazma chemie MeSH
- fotosyntetická reakční centra (proteinové komplexy) chemie metabolismus MeSH
- intracelulární membrány chemie MeSH
- světlosběrné proteinové komplexy chemie metabolismus ultrastruktura MeSH
- transmisní elektronová mikroskopie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Spatial segregation of photosystems in the thylakoid membrane (lateral heterogeneity) observed in plants and in the green algae is usually considered to be absent in photoautotrophs possessing secondary plastids, such as diatoms. Contrary to this assumption, here we show that thylakoid membranes in the chloroplast of a marine diatom, Phaeodactylum tricornutum, contain large areas occupied exclusively by a supercomplex of photosystem I (PSI) and its associated Lhcr antenna. These membrane areas, hundreds of nanometers in size, comprise hundreds of tightly packed PSI-antenna complexes while lacking other components of the photosynthetic electron transport chain. Analyses of the spatial distribution of the PSI-Lhcr complexes have indicated elliptical particles, each 14 × 17 nm in diameter. On larger scales, the red-enhanced illumination exerts a significant effect on the ultrastructure of chloroplasts, creating superstacks of tens of thylakoid membranes.
- MeSH
- chloroplasty metabolismus účinky záření ultrastruktura MeSH
- fotosystém I (proteinový komplex) metabolismus MeSH
- fotosystém II (proteinový komplex) metabolismus MeSH
- multiproteinové komplexy metabolismus ultrastruktura MeSH
- rozsivky metabolismus účinky záření ultrastruktura MeSH
- světlo MeSH
- světlosběrné proteinové komplexy metabolismus MeSH
- transmisní elektronová mikroskopie MeSH
- tylakoidy metabolismus účinky záření ultrastruktura MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A novel chlorophyll a containing pigment-protein complex expressed by cells of Chromera velia adapted to growth under red/far-red illumination [1]. Purification of the complex was achieved by means of anion-exchange chromatography and gel-filtration. The antenna is shown to be an aggregate of ~20kDa proteins of the light-harvesting complex (LHC) family, unstable in the isolated form. The complex possesses an absorption maximum at 705nm at room temperature in addition to the main chlorophyll a maximum at 677nm producing the major emission band at 714nm at room temperature. The far-red absorption is shown to be the property of the isolated aggregate in the intact form and lost upon dissociation. The purified complex was further characterized by circular dichroism spectroscopy and fluorescence spectroscopy. This work thus identified the third different class of antenna complex in C. velia after the recently described FCP-like and LHCr-like antennas. Possible candidates for red antennas are identified in other taxonomic groups, such as eustigmatophytes and the relevance of the present results to other known examples of red-shifted antenna from other organisms is discussed. This work appears to be the first successful isolation of a chlorophyll a-based far-red antenna complex absorbing above 700nm unrelated to LHCI.
- MeSH
- aniontoměniče MeSH
- Apicomplexa metabolismus MeSH
- chlorofyl metabolismus MeSH
- chromatografie iontoměničová MeSH
- cirkulární dichroismus MeSH
- fluorescenční spektrometrie metody MeSH
- gelová chromatografie MeSH
- spektrofotometrie ultrafialová metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The arrangement of core antenna complexes (B808-866-RC) in the cytoplasmic membrane of filamentous phototrophic bacterium Chloroflexus aurantiacus was studied by electron microscopy in cultures from different light conditions. A typical nearest-neighbor center-to-center distance of ~18 nm was found, implying less protein crowding compared to membranes of purple bacteria. A mean RC:chlorosome ratio of 11 was estimated for the occupancy of the membrane directly underneath each chlorosome, based on analysis of chlorosome dimensions and core complex distribution. Also presented are results of single-particle analysis of core complexes embedded in the native membrane.
- MeSH
- buněčná membrána metabolismus ultrastruktura MeSH
- Chloroflexus metabolismus MeSH
- elektronová mikroskopie MeSH
- fotosyntetická reakční centra (proteinové komplexy) metabolismus ultrastruktura MeSH
- fotosyntéza MeSH
- organely metabolismus ultrastruktura MeSH
- Rhodopseudomonas metabolismus MeSH
- světlo MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The authors present a study of the fluorescence and absorbance transients occurring in whole cells of purple nonsulfur bacterium Rhodobacter sphaeroides on the millisecond timescale under pulsed actinic illumination. The fluorescence induction curve is interpreted in terms of combination of effects of redox changes in the reaction center and the membrane potential. The results of this study support the view that the membrane potential act predominantly to increase the fluorescence yield. Advantages of the pulsed actinic illumination for study of the operation of the electron transport chain in vivo are discussed.
- MeSH
- absorpce účinky záření MeSH
- fluorescenční spektrometrie MeSH
- karotenoidy metabolismus MeSH
- kinetika MeSH
- membránové potenciály účinky záření MeSH
- oxidace-redukce účinky záření MeSH
- Rhodobacter sphaeroides cytologie metabolismus účinky záření MeSH
- světlo MeSH
- Publikační typ
- časopisecké články MeSH
