Heterogeneity among photosystem IIα. Isolation of thylakoid membrane vesicles with different functional antennae size of photosystem IIα

doi:10.1016/0005-2728(88)90238-1

Biochimica et Biophysica Acta (BBA) - Bioenergetics

Volume 936, Issue 2, 16 November 1988, Pages 215-221

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Abstract

The functional antenna size of different chloroplast membrane vesicles was determined from ligh-saturation curves of the oxygen evolution using paraphenylbenzoquinone as acceptor. Thylakoids showed a biphasic light-saturation curve indicative of the presence of at least two types of PS II unit, PS II_α and PS II_β. Inside-out thylakoids were isolated by press treatment and phase partition. These vesicles, which contain only PS II_α, were then further fragmented and separated into different classes of vesicles by a sonication-phase partition procedure (Svensson, P. and Albertsson, P.-Å. (1987) in Progress in Photosynthesis Research (Biggins, J., ed.), Vol. II, pp. 281–284, Martinus Nijhoff, Dordrecht). Three fractions enriched in PS II_α were obtained; these differ in the antenna size of PS II_α by up to 20%. It is suggested that the size of the light-harvesting complex of PS II_α from different PS II_α units changes in increments of about 25–50 chlorophyll molecules depending on localization of the units in the grana or different parts of the leaf.

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Cited by (32)

Ultraviolet-B stimulates grana formation in chloroplasts in the African desert plant Dimorphotheca pluvialis
1999, Journal of Photochemistry and Photobiology B: Biology
Chloroplasts isolated from a plant species, Dimorphotheca pluvialis Moench (Asteraceae), grown under visible light without supplementary UV-B radiation, have poor grana development. The dominant components in the membrane organizations are doublets and triplets of stroma lamellae. However, such chloroplasts possess the capacity for electron transfer through the whole photosynthetic chain, based on the measurement of the reduction of ferredoxin. The chloroplasts isolated from D. pluvialis plants grown under visible light in combination with supplementary UV-B radiation have much greater membrane stacking in the thylakoid organization when compared with those grown without supplementary UV-B. The changes in the membrane organization induced by UV-B are also reflected in photochemical activity: oxygen-evolving phoptosystem II (PSII) activity is increased in UV-B chloroplasts compared with chloroplasts grown without UV-B. On this basis, it is concluded that the dynamic relation between the granal stacks and the stroma lamellae in the chloroplasts is closely related to the strategy by which plants are able to cope with a variety of environments and to optimize the photosynthetic machinery for the environmental conditions. The increase in membrane stacking in D. pluvialis, induced by supplementary UV-B radiation during plant growth, suggests that UV-B may be an essential factor for the development of membrane stacking in chloroplasts of higher plants. The mechanism of action of UV-B in this process is discussed.
Antenna protein composition of PS I and PS II in thylakoid sub-domains
1997, Biochimica et Biophysica Acta - Bioenergetics
Spinach thylakoids were separated into grana core, grana margin, and two different stromal lamella fractions in the absence of detergents. The levels of all light-harvesting chlorophyll a/b-binding (LHC) proteins were determined in all fractions, and were normalised to the amount of Photosystem I (PS I) and Photosystem II (PS II) centres. PS Iβ in the stroma lamellae was found to have a full complement of Lhca polypeptides and, probably, one attached LHC II trimer. PS Iα binds additional LHC II trimers, but PS I centres located in the inner parts of the grana stack lack Lhca1 and are depleted in Lhca4. PS IIβ, found in grana margins and stroma lamellae, seems to associate one monomer each of Lhcb4, Lhcb5 and Lhcb6 (CP29, CP26 and CP24, respectively) and one LHC II trimer consisting of two Lhcb1 and one Lhcb3 subunit. PS IIα has additional LHC II trimers (consisting of Lhcb1 and Lhcb2) attached. We also find evidence for the existence of both PS I and PS II centres in the extreme stroma (probably centres being synthesised or repaired), that lack all LHC proteins.
Effects of UVB radiation on light-dependent and light-independent protein phosphorylation in thylakoid proteins
1997, Journal of Photochemistry and Photobiology B: Biology
Protein phosphorylation in thylakoids can be divided into light-dependent and light-independent types. The phosphorylation patterns of these two types of process show about 30 and 40 phosphorylated polypeptides respectively in two-dimensional (2-D) protein separation (molecular mass discrimination in one dimension, isoelectric point (Ip) in the other). It is suggested that each type involves its own signal switch system and kinases responsible for signal transduction from the input domain to the final substrates. A thylakoid protein having a molecular mass of 45 kDa and focusing at Ip 5.0 in the 2-D protein pattern is suggested to be a kinase-like protein and its N-terminal sequence is reported. Freshly prepared spinach thylakoids were exposed to UVB (280–315 nm) radiation emitted from a 4W Philips fluorescent TL12 lamp for 60 min (0.21 W m⁻² plant weighted radiation). The proteins were extracted and subjected to 2-D gel electrophoresis. There was no distinct difference in the 2-D protein patterns between UVB-irradiated thylakoids and non-irradiated thylakoids. The experiment was repeated under conditions of light-dependent protein phosphorylation with [γ-³²P] ATP as phosphate donor. The phosphorylation of a group of polypeptides having molecular masses around 60–65 kDa and isoelectric points around 5–6, thought to include LHCII kinase, was suppressed by UVB pretreatment. The phosphorylation of LHCII polypeptides and other polypeptides was also decreased by UVB treatment. A similar experiment was performed with phosphorylation taking place in the dark after 1 h UVB (or control) treatment. In this case, the UVB treatment had an even greater impact on the phosphorylation pattern. The phosphorylation of a 45 kDa polypeptide, thought to be a kinase-like thylakoid protein active under oxidizing conditions, was almost eliminated, and that of a 60–65 kDa polypeptide group, thought to include LHCII kinase, was strongly suppressed; however, some polypeptides, e.g. 9 kDa and 10 kDa polypeptides focusing at Ip 5.7 and 7.3 respectively, became strongly phosphorylated in UVB-irradiated thylakoids. They may contribute to the protection against UVB radiation. A preliminary model is proposed for the action mechanism of both the enhanced phosphorylated proteins. We believe that altered kinase activity and protein phosphorylation may be important early steps in the impact of UVB radiation on plants.
Differences in UV-B sensitivity between PSII from grana lamellae and stroma lamellae
1996, Journal of Photochemistry and Photobiology B: Biology
UV-B treatment of thylakoid membranes or fractions derived from them accelerates the first rise (F_oF_i phase) of variable fluorescence and slows down the second (F_iF_m) rise. This can be observed for all membrane fractions but to different extents. The grana fraction is affected more by the UV-B treatment than are the intact thylakoids and the stroma lamella fraction. An exogenous electron acceptor, 2,6-dichloro-p-benzoquinone, inhibits both the initial and second rises of the fluorescence induction kinetics including the changes induced by UV-B. However, other electron acceptors, such as 2,5-dimethyl-p-benzoquinone and methyl viologen quench only the second rise of the curve. Oxygen evolution from grana-derived membranes is more sensitive to UV-B treatment than oxygen evolution from stroma lamella-derived membranes.
Size-distributions of antenna and acceptor-pool of Photosystem II
1996, Biochimica et Biophysica Acta - Bioenergetics
A study is described of the heterogeneous kinetics of Photosystem II (PSII) electron transport in osmotically swollen spinach chloroplasts (‘blebs’) on illumination by a series of laser flashes. The existence of a broad distribution of the number of plastoquinone molecules rapidly accessible for reduction by PSII is confirmed by chlorophyll fluorescence yield and electroluminescence measurements as a function of the number of preilluminating flashes. The oscillation of electroluminescence with flash number as a function of flash energy was used to calculate the best-fitting antenna-size distribution of the oxygen evolving PSII. The results suggest that, in blebs at least, electrogenic charge separation leading to electron transport from water to the plastoquinone pool is carried out by PSII β as well as PSII α.
Further characterization of the chloroplast grana margins: the non-detergent preparation of granal Photosystem I cannot reduce ferredoxin in the absence of NADP<sup>+</sup> reduction
1995, BBA - Bioenergetics
The chloroplast grana margins of spinach thylakoids were isolated by sonication and aqueous-two-phase partitioning and their electron transport properties examined. Photosystem II and I electron transport activities were measured and compared to the appressed and non-appressed grana core and stroma lamellae, respectively, as well as to whole thylakoids. The results show that the PS II complexes in the grana margins are of the PS II_β subtype with respect to antenna size, but are Q_B reducing with respect to the acceptor side properties, while the PS I centers in the grana margins have slightly larger antennae as compared to the PS I centers in the stroma lamellae and are more like the PS I_α centers located in the grana domain. The ability to reduce ferredoxin and NADP⁺ was also tested and it was found that the grana margin membrane fraction was unable to reduce ferredoxin, even in the presence of added artificial electron donors. The stroma lamellae and whole thylakoid fractions both reduced ferredoxin at high rates. However, the grana margins could catalyze the reduction of NADP⁺ when supplied with the necessary components (ferredoxin, ferredoxin:NADP⁺ oxidoreductase, and an electron source). The results suggest that the PS I populations located in the margins of the grana domain are functionally different from the PS I centers located in the stroma lamellae. The data support a model whereby the PS I centers in the grana are primarily involved in non-cyclic electron transport, while the PS I centers located in the stroma lamellae are capable of participating in both cyclic and non-cyclic electron transport.

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Regular paperHeterogeneity among photosystem IIα. Isolation of thylakoid membrane vesicles with different functional antennae size of photosystem IIα

Abstract

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

FEBS Lett.

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

Biochim. Biophys. Acta

FEBS Lett.

Biochim. Biophys. Acta

Regular paper
Heterogeneity among photosystem II_α. Isolation of thylakoid membrane vesicles with different functional antennae size of photosystem II_α