Light-harvesting Complex Of Photosystem Research Articles

Photosynthetic Genes of Petunia (Mitchell) Are Differentially Expressed during the Diurnal Cycle

The petunia (Petunia [Mitchell]) chloroplast proteins, the chlorophyll a/b-binding (Cab) proteins, and the small subunit of ribulose bisphosphate carboxylase (RbcS) are encoded by nuclear genes that are expressed in a light-dependent manner. The steady-state concentrations of five cab mRNAs vary with a dramatic circadian rhythm in plants grown under a constant diurnal cycle (10 hours light, 14 hours dark). cab mRNA levels reach their maximum during the light period, but begin to drop prior to the dark period. These RNAs fall to their minimum concentration during the dark period and then begin to increase again in anticipation of the light. Within this general pattern, there are variations in expression among specific classes of cab genes. The light harvesting complex of photosystem II LHCII-type 1 cab mRNAs rise to a well-defined maximum at 2 hours prior to the dark period. All but one of these genes are expressed in anticipation of the light period. The LHCII type 2 cab mRNA and the LHC of photosystem I cab mRNA are expressed at more constant levels throughout the light period. The expression of these genes anticipates the light more than does the expression of the LHCII type 1 genes. The steady state mRNA levels for the petunia rbcS genes show no significant diurnal fluctuation.

Compensatory Alterations in the Photochemical Apparatus of a Photoregulatory, Chlorophyll b-Deficient Mutant of Maize

Characterization of the functional organization of the photochemical apparatus in the light sensitive chlorophyll b-deficient oil yellow-yellow green (OY-YG) mutant of maize (Zea mays) is presented. Spectrophotometric and kinetic analysis revealed substantially lower amounts of the light harvesting complex of photosystem II (LHCII-peripheral) in high light-grown OY-YG thylakoids. However, accumulation of a tightly bound LHCII appears unaffected by the lesion. Changes in photosystem (PS) stoichiometry include lower amounts of PSII with characteristic fast kinetics (PSII(alpha)) and a substantial accumulation of PSII centers with characteristic slow kinetics (PSII(beta)) in the thylakoid membrane of the OY-YG mutant. Thus, PSII(beta) is the dominant photosystem in the mutant chloroplasts. In contrast to wild type, roughly 80% of the mutant PSII(beta) centers are functionally coupled to the plastoquinone pool and are probably localized in the appressed regions of the thylakoid membrane. These centers, designated PSII(beta)-Q(B)-reducing (Q(B) being the secondary electron quinone acceptor of PSII), are clearly distinct from the typical PSII(beta)-Q(B)-nonreducing centers found in the stroma lamellae of wild-type chloroplasts. It is concluded that the observed changes in the stoichiometry of electron-transport complexes reflect the existence of a regulatory mechanism for the adjustment of photosystem stoichiometry in chloroplasts designed to correct any imbalance in light absorption by the two photosystems.

Characterization of a full-length petunia cDNA encoding a polypeptide of the light-harvesting complex associated with photosystem I

We have isolated and characterized a full-length cDNA clone (LHCI-15) which specifies a new chlorophyll-binding protein. This protein is associated with the light-harvesting complex of photosystem I (LHCI). The DNA sequence predicts a precursor protein of 270 amino acids, which shares significant homology with the amino acid sequence of another chlorophyll-binding protein; the chlorophyll a/b-binding (Cab) protein of the photosystem II light-harvesting complex (LHCII). There are two extensive regions of homology (at least 45 residues each) which have approximately 50% amino acid sequence identity. These regions coincide with two of the proposed membrane-spanning alpha helices in the Cab proteins of the LHCII and probably include conserved chlorophyll-binding sites. The LHCI-15 cDNA hybridizes to at least 7 genomic EcoRI DNA fragments, which are very closely related at the nucleotide sequence level.

The origin of the long-wavelength fluorescence emission band (77°K) from photosystem I

Isolated photosystem I (PSI)-110 particles, prepared using a minimal concentration of Triton X-100 [ J. E. Mullet, J. J. Burke, and C. J. Arntzen (1980) Plant Physiol. 65, 814–822 ] and further subjected to short-term solubilization with sodium dodecyl sulfate (SDS), were resolved into four pigment-containing bands on polyacrylamide gel electrophoresis (PAGE). We have identified these in order of increasing electrophoretic mobility as being (a) CPIa, (b) CPI, (c) the light-harvesting complex of photosystem I (LHC-I), and (d) a free pigment-zone. LHC-I had an absorption maximum in the red at 668–669 nm and a shoulder at 650 nm, which was resolved by its first-derivative spectrum to indicate the presence of chlorophyll b. LHC-I exhibited a 77 °K fluorescence emission maximum at 729–730 nm. The 77°K fluorescence emission maxima of CPIa and CPI, excised from the gel, were at 729 and 722 nm, respectively. The LHC-I band, excised from the gel and rerun on dissociating SDS-PAGE, was resolved into two polypeptide doublets of 24-22.5 and 21-20.5 kDa. The CPIa band under similar conditions was resolved into polypeptides of 68, 24, 22.5, 21, 20.5, 19, 15, and 14 kDa; on the contrary, CPI contained only the 68-kDa polypeptide. When intact thylakoids were subjected to “nondenaturing” SDS-PAGE, LHC-I comigrated with an oligomeric form (dimer) of the light-harvesting chlorophyll a/b pigment-protein that preferentially serves photosystem II (LHCP-II). When this combined LHC-I/ LHCP-II pigment-protein band was prepared by SDS-PAGE from isolated stroma lamellae, it exhibited a long-wavelength fluorescence band near 730 nm at 77 °K. When a similar preparation was obtained from sucrose density gradients containing SDS [ J. Argyroudi-Akoyunoglou and H. Thomou (1981) FEBS Lett. 135, 171–181 ], it was found to be enriched in a 21-kDa polypeptide. The data suggest that the 21-kDa polypeptide of LHC-I is the chlorophyll-containing polypeptide responsible for the long-wavelength fluorescence of LHC-I; other polypeptides in the complex (20.5, 22.5, and 24 kDa) presumably bind chlorophyll and also serve an antennae function.

Freeze-fracture analysis of membrane appression and protein segregation in model membranes containing the chlorophyll-protein complexes from chloroplasts.

Cation-induced membrane appression and lateral segregation of chlorophyll-protein complexes have been investigated by freeze-fracture analysis of model membranes containing photosystem 1 and the light-harvesting complex of photosystem 2. In light-harvesting complex proteoliposomes, cations caused extensive membrane adhesion and a segregation of protein into appressed regions. A marked flattening of the appressed membranes, sometimes together with a co-alignment of the particles on the opposing membrane faces, strongly suggests a direct transmembrane attraction between the protein particles. Photosystem 1 membranes were not appressed by cations but some clustering of the particles occurred, together with their exclusion from some regions of the lipid. By incorporating dipalmitoylglycerophosphocholine into the membranes, it is shown that a similar exclusion of the particles can occur due to liquid-crystalline to gel state transitions of the lipids. Proteoliposomes containing both the light-harvesting complex and photosystem 1 displayed cation-induced membrane appression, but only between regions containing the light-harvesting complex. Photosystem 1 was largely confined to unappressed membranes. Destacking occurred at low salt concentrations irrespective of whether photosystem 1 was present, showing that in proteoliposomes, and probably in thylakoids, this process does not require the presence of highly charged complexes from unappressed regions.

Control of chloroplast electron transport by phosphorylation of thylakoid proteins

Several chloroplast membrane proteins can become phosphorylated by a membrane protein kinase. The kinase is activated when the electron carriers (plastoquinone) between photosystem II and photosystem I are reduced. Phosphorylation of the light harvesting complex of photosystem II causes a decrease in energy transferred to photosystem II and increase in the rate of excitation of photosystem I. This process, which is mediated by lateral diffusion of the phosphorylated complex serves to regulate the relative rates of electron transfer through the two photosystems. Phosphorylation may also have an important role in balancing the rate of excitation of photosystem II with the capacity for photosynthetic carbon metabolism and in controlling the functional state of photosystem II.