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1.
LHC II isolated from carnation leaves has been solubilized and resolved by a newly developed, vertical-bed non-denaturing isoelectric focusing in polyacrylamide slab gels to yield three trimeric subcomplexes focusing at pH 4.52, 4.42 and 4.37 (designated a, b and c, respectively), comprising approximately 38%, 24% and 38% of the chlorophyll. The spectroscopic data demonstrated a close similarity among LHC II subcomplexes concerning their chlorophyll content and organization. The most alkaline and the most acidic subcomplex contained the 27 kDa polypeptide of LHC II while the intermediate pI fraction contained both LHC II polypeptides, i.e. 27 kDa and 26 kDa ones associated at 2:1 stoichiometry. The 27 kDa polypeptide could be resolved by denaturing isoelectrofocusing into 10 pI molecular isoforms covering 5.90–4.20 pH range. Three of the isoforms were found in the subcomplexes a and b and eight in the subcomplex c. The 26 kDa polypeptide comprised the unique pI molecular isoform focusing at pH 5.61.Abbreviations CBB G-250
Coomassie Brilliant Blue G-250
- chl
chlorophyll
- DM
n-dodecyl--d-maltoside
- EDTA
ethylendiaminotetraacetic acid
- IEF
isoelectric focusing
- LHC II
the main light-harvesting chlorophyll a/b-protein complex of Photosystem II
- LHCP II
apoprotein of the main light-harvesting chlorophyll a/b-protein complex of Photosystem II
- NP-40
polyethyleneglycol-p-isooctylphenyl ether
- pI
isoelectric point
- OG
octyl--d-glucopyranoside
- PS II
Photosystem II
- SDS-PAGE
sodium dodecylsulphate polyacrylamide gel electrophoresis
- TCA
trichlorooacetic acid 相似文献
2.
The Chl-protein complexes of three maize (Zea mays L.) mutants and one barley (Hordeum vulgare L.) mutant were analyzed using low temperature Chl fluorescence emissions spectroscopy and LDS-polyacrylamide gel electrophoresis. The maize mutants hcf-3, hcf-19, and hcf-114 all exhibited a high Chl fluorescence (hcf) phenotype indicating a disruption of the energy transfer within the photosynthetic apparatus. The mutations in each of these maize mutants affects Photosystem II. The barley mutant analyzed was the well characterized Chl b-less mutant chlorina-f2, which did not exhibit the hcf phenotype. Chlorina-f2 was used because no complete Chl b-less mutant of maize is available. Analysis of hcf-3, hcf-19, and hcf-114 revealed that in the absence of CP43, LHC II can still transfer excitation energy to CP47. These results suggest that in mutant membranes LHC II can interact with CP47 as well as CP43. This functional interaction of LHC II with CP47 may only occur in the absence of CP43, however, it is possible that LHC II is positioned in the thylakoid membranes in a manner which allows association with both CP43 and CP47.Abbreviations
hcf
high chlorophyll fluorescence
- LDS
lithium dodecyl sulfate
- LHC II
light-harvesting complex of Photosystem II
- LHC I
light-harvesting complex of Photosystem I
- CPIa
chlorophyll-protein complex consisting of LHC I and the PS I core complex
- CPI
chlorophyll-protein complex consisting of the PS I core complex
- CP47
47 kDa chlorophyll-protein of the Photosystem II core
- CP43
43 kDa chlorophyll-protein of the Photosystem II core
- CP29
29 kDa chlorophyll-protein of Photosystem II
- CP26
26 kDa chlorophyll-protein of Photosystem II
- CP24
24 kDa chlorophyll-protein of Photosystem II
- fp
free pigments 相似文献
3.
Syetlana V. Manuilskaya Olga I. Volovik Svetlana M. Kochubey 《Photosynthesis research》1995,43(3):225-230
Changes in lipid composition of Photosystem 1 (PS 1) particles isolated from thylakoids phosphorylated under reductive or anaerobic conditions have been studied. Under reductive conditions, there was an increase in monogalactosyldiacylglycerol containing highly saturated fatty acids and phosphatidylglycerol containing transhexadecenoic fatty acid. Under anaerobic conditions, the amount of all lipid classes was increased. As we have shown earlier (S. V. Manuilskaya, O. I. Volovik, A. I. Mikhno, A. I. Polischuk and S. M. Kochubey (1990) Photosynthetica 24: 419–423) these changes were due to a co-migration of some lipid species and light-harvesting chlorophyll a/b complex LHC II from PS 2 to PS 1. These data allow us to conclude that LHC II consists of the lipoproteins containing specific lipids. Different composition of lipids co-migrating with LHC II under various conditions of phosphorylation might be caused by the variety of LHC II subpopulations transferred under each reductive condition.Abbreviations PS 1
Photosystem 1
- PS 2
Photosystem 2
- LHC II
light-harvesting chlorophyll a/b protein complex II
- Chl
chlorophyll
- MGDG
monogalactosyldiacylglycerol
- DGDG
digalactosyldiacylglycerol
- PG
phosphatidylglycerol
- SQDG
sulfoquinovosyldiacylglycerol 相似文献
4.
Assembly of the Light-Harvesting Complexes (LHCs) of Photosystem II (Monomeric LHC IIb Complexes Are Intermediates in the Formation of Oligomeric LHC IIb Complexes) 总被引:7,自引:7,他引:0
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The light-induced assembly of light-harvesting complex (LHC) II has been followed during the biogenesis of the plastid. Seedlings grown in intermittent light (IML) accumulate only small amounts of chlorophyll b. The minor LHC II apoproteins are present; however, the apoprotein levels of the major LHC II complex, LHC IIb, are severely depressed after exposure to IML. The levels of all LHC II apoproteins increase rapidly upon exposure to continuous illumination. The 25-kD, type 3 LHC IIb subunit appears to be more abundant during the early hours of greening in relation to its level in mature thylakoids. The LHC IIb apoproteins are initially associated with pigments to form monomeric pigment-protein complexes. The abundance of monomeric LHC IIb complexes gradually decreases during exposure to continuous light and a concomitant increase occurs in the amount of the trimeric and higher-order oligomeric forms. Pulse-chase experiments verify that labeled LHC IIb monomeric complexes are intermediates in the formation of trimeric and higher-order oligomeric LHC IIb-pigmented complexes. Therefore, the assembly of LHC II occurs via the initial pigmentation of the apoproteins to form monomeric complexes and proceeds in a sequential manner. 相似文献
5.
Pigment protein complexes and the concept of the photosynthetic unit: Chlorophyll complexes and phycobilisomes 总被引:5,自引:0,他引:5
Elisabeth Gantt 《Photosynthesis research》1996,48(1-2):47-53
The photosynthetic unit includes the reaction centers (RC 1 and RC 2) and the light-harvesting complexes which contribute to evolution of one O2 molecule. The light-harvesting complexes, that greatly expand the absorptance capacity of the reactions, have evolved along three principal lines. First, in green plants distinct chlorophyll (Chl) a/b-binding intrinsic membrane complexes are associated with RC 1 and RC 2. The Chl a/b-binding complexes may add about 200 additional chromophores to RC 2. Second, cyanobacteria and red algae have a significant type of antenna (with RC 2) in the form of phycobilisomes. A phycobilisome, depending on the size and phycobiliprotein composition adds from 700 to 2300 light-absorbing chromophores. Red algae also have a sizable Chl a-binding complex associated with RC 1, contributing an additional 70 chromophores. Third, in chromophytes a variety of carotenoid-Chl-complexes are found. Some are found associated with RC 1 where they may greatly enhance the absorptance capacity. Association of complexes with RC 2 has been more difficult to ascertain, but is also expected in chromophytes. The apoprotein framework of the complexes provides specific chromophore attachment sites, which assures a directional energy transfer whithin complexes and between complexes and reaction centers. The major Chl-binding antenna proteins generally have a size of 16–28 kDa, whether of chlorophytes, chromophytes, or rhodophytes. High sequence homology observed in two of three transmembrane regions, and in putative chlorophyll-binding residues, suggests that the complexes are related and probably did not evolve from widely divergent polyphyletic lines.Abbreviations APC
allophycocyanin
- B
phycoerythrin-large bangiophycean phycoerythrin
- Chl
chlorophyll
- LCM
linker polypeptide in phycobilisome to thylakoid
- FCP
fucoxanthin Chl a/c complex
- LHC(s)
Chl-binding light harvesting complex(s)
- LHC I
Chl-binding complex of Photosystem I
- LHC II
Chl-binding complex of Photosystem II
- PC
phycocyanin
- PCP
peridinin Chl-binding complex
- P700
photochemically active Chl a of Photosystem I
- PS I
Photosystem I
- PS II
Photosystem II
- RC 1
reaction center core of PS I
- RC 2
reaction center core of PS II
- R
phycoerythrin-large rhodophycean phycoerythrin
- sPCP
soluble peridinin Chl-binding complex 相似文献
6.
T. Bei-Paraskevopoulou R. Anastassiou J. Argyroudi-Akoyunoglou 《Photosynthesis research》1995,44(1-2):93-106
The appearance of the light harvesting II (LHC II) protein in etiolated bean leaves, as monitored by immunodetection in LDS-solubilized leaf protein extracts, is under phytochrome control. A single red light pulse induces accumulation of the protein, in leaves kept in the dark thereafter, which follows circadian oscillations similar to those earlier found for Lhcb mRNA (Tavladoraki et al. (1989) Plant Physiol 90: 665–672). These oscillations are closely followed by oscillations in the capacity of the leaf to form Chlorophyll (Chl) in the light, suggesting that the synthesis of the LHC II protein and its chromophore are in close coordination. Experiments with levulinic acid showed that PChl(ide) resynthesis does not affect the LHC II level nor its oscillations, but new Chl a synthesis affects LHC II stabilization in thylakoids, implicating a proteolytic mechanism. A proteolytic activity against exogenously added LHC II was detected in thylakoids of etiolated bean leaves, which was enhanced by the light pulse. The activity, also under phytochrome control, was found to follow circadian oscillations in verse to those in the stabilization of LHC II protein in thylakoids. Such a proteolytic mechanism therefore, may account for the circadian changes observed in LHC II protein level, being implicated in pigment-protein complex assembly/stabilization during thylakoid biogenesis.Abbreviations Chl
chlorophyll
- CL
continuous light
- D
dark
- FR
far-red light
- LA
levulinic acid
- LHC II
light-harvesting complex serving Photosystem II
- PChl(ide)
protochlorophyllide
- PCR
protochlorophyllide oxidoreductase
- R
red light 相似文献
7.
Despite the total loss of Photosystem II activity, thylakoids isolated from the green nuclear maize mutant hcf1-3 contain normal amounts of the light-harvesting chlorophyll pigment-protein complex (LHC). We interpret the spectroscopic and ultrastructural characteristics of these thylakoids to indicate that the LHC present in these membranes is not associated with Photosystem II reaction centers and thus exists in a ‘free’ state within the thylakoid membrane. In contrast, the LHC found in wild-type maize thylakoids shows the usual functional association with Photosystem II reaction centers. Several lines of evidence suggest that the free LHC found in thylakoids isolated from hcf1-3 is able to mediate cation-dependent changes in both thylakoid appression and energy distribution between the photosystems: (1) Thylakoids isolated from hcf1-3 and wild-type seedlings exhibit a similar Mg2+-dependent increase in the short/long wavelength fluorescence emission peak ratio at 77 K. This Mg2+ effect is lost following incubation of thylakoids isolated from either source with low concentrations of trypsin. Such treatment results in the partial proteolysis of the LHC in both membrane types. (2) Thylakoids isolated from both hcf1-3 and wild-type seedlings show a similar Mg2+ dependence for the enhancement of the maximal yield of room temperature fluorescence and light scattering; both Mg2+ effects are abolished by brief incubation of the thylakoids with low concentrations of trypsin (3) Mg2+ acts to reduce the relative quantum efficiency of Photosystem I-dependent electron transport at limiting 650 nm light in thylakoids isolated from hcf1-3. (4) The pattern of digitonin fractionation of thylakoid membranes, which is dependent upon structural membrane interactions and upon LHC in the thylakoids, is similar in thylakoids isolated from both hcf1-3 and wild-type seedlings. We conclude that the surface-exposed segment of the LHC, but not the LHC-Photosystem II core association, is necessary for the cation-dependent changes in both thylakoid appression and energy distribution between the two photosystems, and that the LHC itself is able to transfer excitation energy directly to Photosystem I in a Mg2+-dependent fashion in the absence of Photosystem II reaction centers. The latter phenomenon is equivalent to a cation-induced change in the absorptive cross-section of Photosystem I. 相似文献
8.
Organization of the Light-Harvesting Complex of Photosystem I and Its Assembly during Plastid Development 总被引:7,自引:6,他引:1
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Photosystem I (PSI) holocomplexes were fractionated to study the organization of the light-harvesting complex I (LHC I) pigment-proteins in barley (Hordeum vulgare) plastids. LHC Ia and LHC Ib can be isolated as oligomeric, presumably trimeric, pigment-protein complexes. The LHC Ia oligomeric complex contains both the 24- and the 21.5-kD apoproteins encoded by the Lhca3 and Lhca2 genes and is slightly larger than the oligomeric LHC Ib complex containing the Lhca1 and Lhca4 gene products of 21 and 20 kD. The synthesis and assembly of LHC I during light-driven development of intermittent light-grown plants occurs rapidly upon exposure to continuous illumination. Complete PSI complexes are detected by nondenaturing Deriphat (disodium N-dodecyl-[beta]-iminodipropionate-160)-PAGE after 2 h of illumination, and their appearance correlates with that of the 730- to 740-nm emission characteristic of assembled LHC I. However, the majority of the newly synthesized LHC I apoproteins are present as monomeric complexes in the thylakoids during the early hours of greening. We propose that during development of the protochloroplast the LHC I apoproteins are first assembled into monomeric pigmented complexes that then aggregate into trimers before becoming attached to the pre-existing core complex to form a complete PSI holocomplex. 相似文献
9.
The precursor to the nuclear-coded 17 kDa early light-inducible protein (ELIP) of pea has been transported into isolated intact chloroplasts. The location of the mature protein in the thylakoid membranes was investigated after using cleavable crosslinkers such as DSP and SAND in conjunction with immuno-fractionation methods and by application of mild detergent fractionation. We show that ELIP is integrated into the membranes via the unstacked stroma thylakoids. After isolation of protein complexes by solubilization of membranes with Triton X-100 and sucrose density-gradient centrifugation the crosslinked ELIP comigrates with the PS II core complex. Using SAND we identified ELIP as a 41–51 kDa crosslinked product while with DSP four products of 80 kDa, 70 kDa, 50–42 kDa and 23–21 kDa were found. The immunoprecipitation data suggested that the D1-protein of the PS II complex is one of the ELIP partners in crosslinked products.Abbreviations chl
chlorophyll
- D1
herbicide-binding protein
- DSP
dithiobis-(succinimidylpropionate)
- ELIP
early light-inducible protein
- LHC I and LHC II
light-harvesting chlorophyll a/b complex associated with photosystem I or II
- PAGE
polyacrylamide gel electrophoresis
- poly(A)-rich RNA
polyadenyd mRNA
- PS I and PS II
photosystems I and II
- SAND
sulfosuccinimidyl 2-(m-azido-o-nitro-benzamido)-ethyl-1,3-dithiopropionate
- Triton X-100
octylphenoxypolyethoxyethanol 相似文献
10.
The transverse orientation of the light-harvesting chlorophyll a/b protein complex of Photosystem II (LHC II) in the thylakoid membrane of pea was investigated using surface radioiodination with Iodo-GenTM. The labelling effects on LHC II of four different membrane preparations were compared. One preparation was oriented right-side-out (intact thylakoids); two of them had an inside-out orientation exposing the lumenal surface (inside-out vesicles; PS II particles) and one had both sides of the membrane exposed (mechanically damaged thylakoids). It was found that LHC II could be iodinated only in membrane preparations with an exposed lumenal surface. Isolated apoproteins were chemically cleaved. Fragments analysis revealed a tyrosine residue located eight amino acids from the C-terminus as the single iodination site. It is concluded that the C-terminus of LHC II points towards the lumental side of the thylakoid. Differences in the labelling behaviour of the LHC apoproteins could be assigned to a heterogeneity in the C-terminal region in which the tyrosine residue is replaced by phenylalanine. 相似文献
11.
Phosphatase activities in spinach thylakoid membranes-effectors, regulation and location 总被引:1,自引:0,他引:1
The dephosphorylation of seven phosphoproteins associated with Photosystem II or its chlorophyll a/b antenna in spinach thylakoids, was characterised. The rates were found to fall into two distinct groups. One, rapidly dephosphorylated, consisted of the two subunits (25 and 27 kD) of the major light harvesting complex of Photosystem II (LHC II) and a 12 kD polypeptide of unknown identity. A marked correlation between the dephosphorylation of these three phosphoproteins, strongly suggested that they were all dephosphorylated by the same enzyme. Within this group, the 25 kD subunit was consistently dephosphorylated most rapidly, probably reflecting its exclusive location in the peripheral pool of LHC II. The other group, only slowly dephosphorylated, included several PS II proteins such as the D1 and D2 reaction centre proteins, the chlorophyll-a binding protein CP43 and the 9 kD PS II-H phosphoprotein. No dephosphorylation was observed in either of the two groups in the absence of Mg2+-ions. Dephosphorylation of the two LHC II subunits took place in both grana and stroma-exposed regions of the thylakoid membrane. However, deposphorylation in the latter region was significantly more rapid, indicating a preferential dephosphorylation of the peripheral (or mobile) LHC II. Dephosphorylation of LHC II was found to be markedly affected by the redox state of thiol-groups, which may suggest a possible regulation of LHC II dephosphorylation involving the ferredoxin-thioredoxin system.Abbreviations CP 43
43 kD chlorophyll a- binding protein
- D1 and D2
reaction centre proteins of PS II
- LHC II
light-harvesting complex of PS II
- LHC II-25
25 kD subunit of LHC II
- LHC II-27
27 kD subunit of LHC II
- NEM
N-ethylmaleimide
- PP2C
protein phosphatase 2C
- PS II-H
psb H gene product 相似文献
12.
Triton X-100, a detergent commonly used to solubilize higher plant thylakoid membranes, was found to be deleterious to Dunaliella LHC II. It disrupted the transfer of excitation energy from chlorophyll b to chlorophyll a. Based on analysis of pigments and immunoassays of LHC II apoproteins from sucrose density gradient fractions, Triton X-100 caused aggregation of the complex, but apparently did not remove chlorophyll b from the apoprotein. Following solubilization with Triton X-100 only CPI could be resolved by electrophoresis. In contrast, solubilization of Dunaliella thylakoids with octyl--D-glucopyranoside preserved energy transfer from chlorophyll b to chlorophyll a. This detergent also effectively prevented aggregation on sucrose gradients and preserved CPI oligomers, as well as LHCP1 and LHCP3 on non-denaturing gels. Solubilization with Deriphat gave similar results. We propose that room temperature fluorescence excitation and emission spectroscopy be used in conjunction with other biophysical and biochemical probes to establish the effects of detergents on the integrity of light harvesting chlorophyll protein complexes. Methods used here may be applicable to other chlorophytes which prove refractory to protocols developed for higher plants.Abbreviations LHC II
light harvesting chlorophyll protein complex associated with photosystem II
- LHCP1 and LHCP3
monomeric and oligomeric forms of LHC II, respectively, observed on non-denaturing gels
- LiDS
lithium dodecylsulphate
- PMSF
phenylmethylsulfonyl fluoride 相似文献
13.
Reversible changes in the room temperature fluorescence quenching at 685 nm and light scattering level at 577 nm, indicating about 15% of granal unstacking, induced by high temperature treatment (40°C, for 5 min) of pea chloroplasts were shown. Analysis of the low temperature excitation fluorescence spectra of the 735 nm Photosystem 1 (PS 1) band (F735), in the 635–725 nm region, has revealed the involvement of light-harvesting (LHC 2, maxima at 650 and 676 nm) and the proximal Photosystem 2 antenna (maxima 668, 687 nm) in heat-induced enhancement of the PS 1 long wavelength antenna absorption cross-section. It was found that the two PS 1 sub-chloroplast preparations, achieved by the digitonin method, possessed different characteristics of this enhancement. For the heavier fraction (100 000 g) the additional absorption cross-section was formed mostly at the expense of PS 2 antennas (apparently spillover), but for the lighter PS 1 fraction (145 000 g) the changes have indicated an -transfer mechanism, i.e., participation of only LHC 2 in the energy transfer towards PS 1. This may indicate the heterogeneous character of the temperature-induced energy redistribution across the PS 1-containing chloroplast membrane compartments. The model of heat-induced changes in the pigment-protein complex arrangement is discussed in terms of domain organisation of the thylakoid membrane.Abbreviations Chl a/b
ratio between chlorophyll a and chlorophyll b concentrations
- CP43 and CP47
proximal Photosystem 2 antenna complexes
- D1/D2 complex
Photosystem 2 reaction centre complex
- EDTA
ethylenediaminetetraacetic acid
- F685 and F696
Photosystem 2 low temperature fluorescence bands
- F735
Photosystem 1 low temperature fluorescence band
- Fp
free pigment band in green gel electrophoresis
- LHC 2
light-harvesting chlorophyll a/b complex
- LHCP I, II and III
light-harvesting bands in green gel electrophoresis
- Cp1 and Cpa
bands in green gel electrophoresis which are associated with Photosystem 1 and 2 reaction centre complexes with internal antennas
- P700
Photosystem 1 reaction centre
- PPC
pigment-protein complex
- PS 1 and
Photosystem 1 alpha and Photosystem 1 beta
- PS 2 and
Photosystem 2 alpha and Photosystem 2 beta
- RC
reaction centre
- SDS-PAGE
sodiumdodecylsulphate-polyacrylamide gel electrophoresis
- St1-St2
state-1-state-2 transitions 相似文献
14.
The major light-harvesting chlorophyll a/b-protein (LHCP) of higher plant chloroplasts is a nuclearencoded, integral thylakoid membrane protein that binds photosynthetic pigments and occurs in situ in an oligomeric form. We have previously examined structural and functional domains of the mature apoprotein by use of mutant LHCPs and in vitro assays for uptake and insertion. Results presented here demonstrate the effects of several mutations in the amino terminal domain of the mature apoprotein. Deletion of amino acid residues 12–58 greatly affected import into chloroplasts, while deletion or alteration of the hydrophobic region E65VIHARWAM73 led to rapid degradation of the mutant LHCP. We suggest that this amino-proximal region is essential for the stability of the LHCP and its ability to integrate into the thylakoid membranes. A structural/functional relationship of this region to a previously examined hydrophobic carboxy-proximal domain [Kohorn and Tobin (1989), The Plant Cell 1, 159–166] is proposed.Abbreviations BSA
bovine serum albumin faction V
- ELIPs
early light-inducible proteins
- Hepes
4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- LHCP
light-harvesting chlorophyll a/b-protein
- LHC IIb
light-harvesting complex associated with Photosystem II
- pLHCP
precursor to LHCP
- Rubisco
ribulose 1,5-biphosphate carboxylase-oxygenase
- SDS-PAGE
sodium dodecyl sulfate-poly-acrylamide gel electrophoresis 相似文献
15.
Summary Irradiation of the principal photosystem II light-harvesting chlorophyll-protein antenna complex, LHC II, with high light intensities brings about a pronounced quenching of the chlorophyll fluorescence. Illumination of isolated thylakoids with high light intensities generates the formation of quenching centres within LHC II in vivo, as demonstrated by fluorescence excitation spectroscopy. In the isolated complex it is demonstrated that the light-induced fluorescence quenching: a) shows a partial, biphasic reversibility in the dark; b) is approximately proportional to the light intensity; c) is almost independent of temperature in the range 0–30°C; d) is substantially insensitive to protein modifying reagents and treatments; e) occurs in the absence of oxygen. A possible physiological importance of the phenomenon is discussed in terms of a mechanism capable of dissipating excess excitation energy within the photosystem II antenna.Abbreviations chla
chlorophyll a
- chlb
chlorophyll b
- F0
fluorescence yield with reaction centers open
- Fm
fluorescence yield with reaction centres closed
- Fi
fluorescence at the plateau level of the fast induction phase
- LHC II
light-harvesting chlorophyll a/b protein complex II
- PS II
photosystem II
- PSI
photosystem I
- Tricine
N-[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine 相似文献
16.
Jacqueline Olive Francis-André Wollman Pierre Bennoun Michel Recouvreur 《Archives of biochemistry and biophysics》1981,208(2):456-467
The ultrastructure of the thylakoid membranes of Chlamydomonas reinhardtii was investigated using cell cultures grown under light intensities of 200 and 4000 lx, respectively. A significant difference in the size distribution of the exoplasmic fracture face (EF) particles appears upon Mg2+ treatment of broken cell preparations from the two light growth conditions. Particles larger than 150 Å are seen at 4000 lx only. However neither the absorption spectra of chlorophyll at 77 °K, nor the chlorophyll a/chlorophyll b ratios differ in the two cell batches. In addition, the polypeptide composition of the thylakoid membranes and the Mg2+ effect (spillover) on the photochemical rate of Photosystem II are the same in both conditions. We conclude that the partition coefficient between the two fracture faces of light-harvesting complex-containing particles is variable. It depends on Mg2+ ion concentration in the incubating medium of the membranes and on the light growth conditions of the cell cultures. Our results suggest that 60- to 80-Å protoplasmic fracture face (PF) particles containing the light-harvesting complexes can aggregate either in larger PF particles (100–120 Å) or in EF particles larger than 120 Å which also contain the Photosystem II centers. That some light-harvesting complexes are located on the PF faces is confirmed by the analysis of the BF4 mutant of C. reinhardtii lacking in chlorophyll-protein complex II. The PF faces of the BF4 thylakoids display a reduced number of particles as compared to that in the wild type. 相似文献
17.
In recent years major progress has been made in describing the gene families that encode the polypeptides of the light-harvesting antenna system of photosystem II (PSII). At the same time, advances in the biochemical characterization of these antennae have been hampered by the high degree of similarity between the apoproteins. To help interpret the molecular results, we have re-examined the composition, the assembly and the phosphorylation patterns of the light-harvesting antenna of PSII (LHCII) in the green alga Chlamydomonas reinhardtii Dang, using a non-Tris SDS-PAGE system capable of resolving polypeptides that differ by as little as 200 daltons. Research to date has suggested that in C. reinhardtii the LHCII comprises just four polypeptides (p11, p13, p16 and p17), and CP29 and CP26 just one polypeptide each (p9 and p10, respectively), i.e. a total of six polypeptides. We report here that these antenna systems contain at least 15 polypeptides, 10 associated with LHCII, 3 with CP29, and 2 with CP26. All of these polypeptides have been positively identified by means of appropriate antibodies. We also demonstrate substantial heterogeneity to the pattern of in-vitro phosphorylation, with major differences found among members of closely spaced and immunologically related polypeptides. Most intriguing is the fact that the polypeptides that cross-react with the anti-type 2 LHCII antibodies of higher plants (p16, and to a lesser extent p11) are not phosphorylated, whereas in higher plants these are the most highly phosphorylated polypeptides. Also, unlike in higher plants, CP29 is heavily phosphorylated. Phosphorylation does not appear to have any effect on the mobility of polypeptides on fully denaturing SDS-PAGE gels. To learn more about the accumulation and organization of the light-harvesting polypeptides, we have also investigated a chlorophyll b-less mutant, cbn1-48. The LHCII is almost completely lost in this mutant, along with at least some LHCI. But the accumulation of CP29 and CP26 and their binding to PSII core complexes, is relatively unaffected. As expected, the loss of antenna polypeptides is accompanied by a reduction of the size of large reaction-center complexes. Following in-vitro phosphorylation the number of phosphorylated proteins is greatly increased in the mutant thylakoids compared to wildtype thylakoids. We present a model of the PSII antenna system to account for the new polypeptide complexity we have demonstrated.This work was supported by National Institute of Health grant GM22912 to L.A.S. We would like to thank Anastasios Melis for helpful discussions. 相似文献
18.
19.
Xanthophyll Cycle Pigment Localization and Dynamics during Exposure to Low Temperatures and Light Stress in Vinca major 总被引:4,自引:0,他引:4
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Amy S. Verhoeven William W. Adams III Barbara Demmig-Adams Roberta Croce Roberto Bassi 《Plant physiology》1999,120(3):727-738
The distribution of xanthophyll cycle pigments (violaxanthin plus antheraxanthin plus zeaxanthin [VAZ]) among photosynthetic pigment-protein complexes was examined in Vinca major before, during, and subsequent to a photoinhibitory treatment at low temperature. Four pigment-protein complexes were isolated: the core of photosystem (PS) II, the major light-harvesting complex (LHC) protein of PSII (LHCII), the minor light-harvesting proteins (CPs) of PSII (CP29, CP26, and CP24), and PSI with its LHC proteins (PSI-LHCI). In isolated thylakoids 80% of VAZ was bound to protein independently of the de-epoxidation state and was found in all complexes. Plants grown outside in natural sunlight had higher levels of VAZ (expressed per chlorophyll), compared with plants grown in low light in the laboratory, and the additional VAZ was mainly bound to the major LHCII complex, apparently in an acid-labile site. The extent of de-epoxidation of VAZ in high light and the rate of reconversion of Z plus A to V following 2.5 h of recovery were greatest in the free-pigment fraction and varied among the pigment-protein complexes. Photoinhibition caused increases in VAZ, particularly in low-light-acclimated leaves. The data suggest that the photoinhibitory treatment caused an enrichment in VAZ bound to the minor CPs caused by de novo synthesis of the pigments and/or a redistribution of VAZ from the major LHCII complex. 相似文献
20.
Using absorption and fluorescence experiments at low temperature with polarized light on oriented samples, the orientation of PS-I-related pigments, both in green plants and in Chlamydomonas reinhardtii, has been investigated on isolated pigment-protein complexes and intact thylakoids. The following observations have been made. (i) The isolation procedure of PS I110, PS I65, LHC I and CP0) particles from pea and C. reinhardtii do not alter significantly the intrinsic orientation of the pigments inside the complexes; (ii) Chl b is a structural component of PS I, linked to the peripheral antenna, with an orientation with respect to the thylakoid plane different from that observed in the main light-harvesting complex (iii) PS I65 (i.e., ‘core’ PS I) of pea and C. reinhardtii contains identical chromophores having the same orientation with respect to the geometrical longest axis (axes) of the complexes. (iv) LHC I and CP0 (i.e., PS I ‘peripheral antenna’) of pea and C. reinhardtii have identical oriented chromophores, except that a long-wavelength component with a high anisotropy is only present in green plants. This set of pigments, which absorbs at 705–725 nm, has the same orientation as the dipoles emitting F735 and also as the QY transition of P-700. (v) All the long-wavelength fluorescence properties of the various studied membranes are explained by these data on isolated PS I complexes: wild-type C. reinhardtii and Chl-b-less barely fluoresce from the core pigments, while a CP1 deficient mutant of C. reinhardtii and wild-type barley fluoresce from the antenna pigments. 相似文献