Isolation and characterization of the chlorophyll a/b protein complex CP29 from spinach

Henrysson, Tomas; Schröder, Wolfgang P.; Spangfort, Michael D.; �kerlund, Hans-Erik

doi:10.1016/s0005-2728(89)80084-2

Cited by 54 publications

(39 citation statements)

References 35 publications

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“…There is up to 90% sequence identity between the spinach protein and the others, confirming that this protein is LhcbS. The spinach 30-kDa protein (from fraction 8) showed 100% identity with the known sequence for spinach Lhcb4, the LHCIIa apoprotein (Henrysson et al, 1989), and strong similarity with the predicted sequence for barley Lhcb4 (Morishige and Thornber, 1992), confirming that the major complex in fraction 8 is indeed LHCIIa. Binding to LH.CIIa, LHCIIb and LHCIIc isolated by IEF by 50 pM (cHxN),'~C under similar conditions to those used previously confirmed that the stoichiometry of binding to LHCIIb was low compared with that to LHCIIa and LHCIIc (Fig.…”

Section: (Chxn)c Labelling Of Purified Lhcii Polypeptides In Vitromentioning

confidence: 74%

“…SDShrea PAGE was carried out as previously described (Walters and Horton, 1995). Electrophoresis was carried out overnight at room temperature (approximately 15 "C) ; these conditions allow clear separation of LHCIIa and LHCIIb apoproteins (Henrysson et al, 1989).…”

Section: Electrophoresismentioning

confidence: 99%

“…N-terminal amino-acid sequences of tryptic fragments of LHCII apoproteins. Amino-acid sequence data from fragments of the proteins in IEF fraction 6 (26 kDa) and fraction 8 (30 ma), aligned with the corresponding sequences from LHCIIc from Scots Pine (Jansson, 1992), tomato (Schwartz et al, 1991) and barley (SGrensen et al, 1992), and from LHCIIa from spinach (Henrysson et al, 1989) and barley (Morishige and Thornber, 1992). Aminoacids whose determination was uncertain are shown in parenthesis.…”

Section: (Chxn)c Labelling Of Photosystem I1 Proteinsmentioning

confidence: 99%

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Higher Plant Light‐Harvesting Complexes LHCIIa and LHCIIc are Bound by Dicyclohexylcarbodiimide During Inhibition of Energy Dissipation

Walters

Ruban

Horton

1994

European Journal of Biochemistry

126

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We have investigated the binding to proteins of the photosynthetic apparatus of the carboxymodifying agent dicyclohexylcarbodiimide, (cHxN),C ; this inhibits the protective dissipation of excess absorbed light energy (qE) by the light-harvesting apparatus of photosystem I1 (LHCII), suggesting that carboxyl amino-acid side chains within hydrophobic protein domains may be involved in qE. (cHxN),I4C was used to label thylakoids and photosystem I1 particles, so as to identify proteins which may be involved in the detection of lumen pH during qE induction. Of six thylakoid proteins labelled with (cHxN),C under conditions where qE is efficiently induced, three are associated with photosystem I, and none with the bulk LHCII. PSII-associated label is bound to three minor components of LHCII, identified as LHCIIa (two species) and LHCIIc, as shown by protein sequencing of tryptic fragments of purified complexes. pH titration of qE formation and protein labelling in coupled thylakoids showed that both qE and labelling of LHCIIa increased at pH 7-8.In higher plants, chlorophyll and carotenoid molecules are bound to specific proteins to form light-harvesting complexes (LHCI, LHCII), which efficiently capturc and deliver excitation energy to photosystem I (PSI) and photosystem I1 (PSII) reaction centres. Four different types of LHCII (LHCIIa-d), which differ in their polypeptide and pigment content, have been identified (Peter and Thornber, 1991). The polypeptides comprising LHCII are encoded by members of the Lhc gene family; Lhcbl, Lhcb2 and Lhcb3 encode 28-, 27-and 25-kDa polypeptides, respectively, comprising LHCIIb; Lhcb4 encodes the 30-31-kDa protein of LHCIIa; Lhcb.5 encodes the LHCIIc 26-kDa protein; the LHCIId 24-kDa protein is encoded by Lhcbd . LHCIIb binds up to 65% of PSII chlorophyll and is often referred to as the bulk light-harvesting complex; LHCIIa, c and d are minor complexes, each accounting for 3-5% (Peter and Thornber, 1991 ;Dainese and Bassi, 1991 ;.When the light absorbed by LHCII is greater than that capable of being used in photosynthesis, the photosynthetic apparatus is protected from the damaging effects of overexcitation by the rapid induction of a process of non-radiative energy dissipation known as qE Demmig-Adams, 1990;Horton and Ruban, 1992). Measurement of radiationless dissipation of energy via non-photochemical

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Section: (Chxn)c Labelling Of Purified Lhcii Polypeptides In Vitromentioning

confidence: 74%

Section: Electrophoresismentioning

confidence: 99%

Section: (Chxn)c Labelling Of Photosystem I1 Proteinsmentioning

confidence: 99%

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Higher Plant Light‐Harvesting Complexes LHCIIa and LHCIIc are Bound by Dicyclohexylcarbodiimide During Inhibition of Energy Dissipation

Walters

Ruban

Horton

1994

European Journal of Biochemistry

126

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“…The first panel is the silver-stained pattern, and the other panels are immunoblots of similar gels. The nine polypeptides labeled in this figure have been assigned on the basis of the antibody binding described below and on published characterizations (4,5,12,16). The second panel shows immunoreactivity with an antiserum prepared against purified spinach LHCII (14).…”

Section: Separation Of Cp Complexes By Fluid Phase Iefmentioning

confidence: 99%

“…CP26 also has a higher Chl a/b ratio than LHCII but generally comigrates with LHCII in both native and fully denaturing SDS-PAGE systems (4,27). Coupled with the difficulty in separating CP29 from the LHCII apoproteins in some species (particularly in spinach [16,17]), this has contributed to confusion in the literature as to the identities of these two complexes. This issue has recently been settled by the comparison of partial protein sequence data for CP29 and CP26 in barley (Hordeum vulgare) with a full-length gene sequence for CP29 (26), confirming that CP29 and CP26 are distinct complexes, both from each other and from LHCII.…”

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confidence: 99%

Biochemical Characterization of Photosystem II Antenna Polypeptides in Grana and Stroma Membranes of Spinach

Allen¹,

Staehelin²

1992

Plant Physiol.

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The photosystem (PS) II antenna system comprises several biochemically and spectroscopically distinct complexes, including light-harvesting complex 11 (LHCII), chlorophyll-protein complex (CP) 29, CP26, and CP24. LHCII, the most abundant of these, is both structurally and functionally diverse. The photosynthetic apparatus is laterally segregated within the thylakoid membrane into PSI-rich and PSII-rich domains, and the distribution of antenna complexes between these domains has implications for antenna function. We report a detailed analysis of the differences in the polypeptide composition of LHCII, CP29, and CP26 complexes associated with grana and stroma thylakoid fractions from spinach (Spinacia oleracea L.), making use of a very high-resolution denaturing gel system, coupled with immunoblots using monospecific antibodies to identify specific antenna components. We first show that the polypeptide composition of the PSII antenna system is more complex than previously thought. We resolved at least five type I LHCII apoproteins and two to three type 11 LHCII apoproteins. We also resolved at least two apoproteins each for CP29 and CP26. In state 1-adapted grana and stroma thylakoid membranes, the spectrum of LHCII apoproteins is surprisingly similar. However, in addition to overall quantitative differences, we saw subtle but reproducible qualitative differences in the spectrum of LHCII apoproteins in grana and stroma membrane domains, including two forms of the major type 11 apoprotein. The implications of these findings for models of PSII antenna function in spinach are discussed.Most of the Chl in the thylakoid membrane of higher plants and green algae is bound by specialized antenna complexes associated with either PSI or PSII (5,15,33 Chl. The CP29 complex has a Chl a/b ratio of 2.2 to 2.5 (5, 27) and has an apoprotein several kD larger than those of the LHCII complex. CP26 also has a higher Chl a/b ratio than LHCII but generally comigrates with LHCII in both native and fully denaturing SDS-PAGE systems (4, 27). Coupled with the difficulty in separating CP29 from the LHCII apoproteins in some species (particularly in spinach [16,17]), this has contributed to confusion in the literature as to the identities of these two complexes. This issue has recently been settled by the comparison of partial protein sequence data for CP29 and CP26 in barley (Hordeum vulgare) with a full-length gene sequence for CP29 (26), confirming that CP29 and CP26 are distinct complexes, both from each other and from LHCII.In this paper we will use the terms CP26 and CP29 as defined in ref. 4. Little is known for certain about the physical arrangement of the PSII antenna system, but the isolation of 02-evolving complexes lacking LHCII but still containing CP29 suggests that CP29 is more closely associated with the reaction center (9). The gel systems used in ref. 9 did not distinguish between CP29 and CP26; therefore, no conclusions could be drawn about the relationship of CP26 to the reaction center.

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Towards the Atomic Resolution Structure of Light Harvesting Antennae

Ruban

2012

The Photosynthetic Membrane

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Isolation and characterization of the chlorophyll a/b protein complex CP29 from spinach

Cited by 54 publications

References 35 publications

Higher Plant Light‐Harvesting Complexes LHCIIa and LHCIIc are Bound by Dicyclohexylcarbodiimide During Inhibition of Energy Dissipation

Higher Plant Light‐Harvesting Complexes LHCIIa and LHCIIc are Bound by Dicyclohexylcarbodiimide During Inhibition of Energy Dissipation

Biochemical Characterization of Photosystem II Antenna Polypeptides in Grana and Stroma Membranes of Spinach

Towards the Atomic Resolution Structure of Light Harvesting Antennae

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