PIGMENT COMPLEXES OF LIGHT‐HARVESTING CHLOROPHYLL <i>a/b</i> BINDING PROTEIN ARE STABILIZED BY A SEGMENT IN THE CARBOXYTERMINAL HYDROPHILIC DOMAIN OF THE PROTEIN*

Paulsen, Harald; Kuttkat, Andrea

doi:10.1111/j.1751-1097.1993.tb02269.x

Cited by 22 publications

(28 citation statements)

References 12 publications

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“…In contrast to former studies on the reconstitution of LHCII monomers which took place on an analytical scale (Paulsen and Hobe, 1992;Paulsen and Kuttkat, 1993) (Shipley et al, 1973) thereby preventing the formation of a homogenous population of lipid vesicles of known concentration. Eluted fractions which did not contain MGDG were pooled, evaporated to dryness and resuspended in buffer T at a concentration of 10 mg/ml by sonification.…”

Section: Methodsmentioning

confidence: 78%

Trimerization and crystallization of reconstituted light-harvesting chlorophyll a/b complex.

et al. 1994

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The major light-harvesting complex (LHCII) of photosystem II, the most abundant chlorophyll-containing complex in higher plants, is organized in trimers. In this paper we show that the trimerization of LHCII occurs spontaneously and is dependent on the presence of lipids. LHCII monomers were reconstituted from the purified apoprotein (LHCP), overexpressed in Escherichia coli, and pigments, purified from chloroplast membranes. These synthetic LHCII monomers trimerize in vitro in the presence of a lipid fraction isolated from pea thylakoids. The reconstituted LHCII trimers are very similar to native LHCII trimers in that they are stable in the presence of mild detergents and can be isolated by partially denaturing gel electrophoresis or by centrifugation in sucrose density gradients. Moreover, both native and reconstituted LHCII trimers exhibit signals in circular dichroism in the visible range that are not seen in native or reconstituted LHCII monomers, indicating that trimer formation either establishes additional pigment-pigment interactions or alters pre-existing interactions. Reconstituted LHCII trimers readily form two-dimensional crystals that appear to be identical to crystals of the native complex.

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Section: Methodsmentioning

confidence: 78%

Trimerization and crystallization of reconstituted light-harvesting chlorophyll a/b complex.

et al. 1994

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“…The precursor pLHCP and its derivatives were expressed from the plasmid XLHCP-2 (Paulsen et al, 1990) containing the coding region of the cab gene AB80 (Ihb 1*2, according to Jansson el al., 1992) from pea (Pisurn sativurn) (Cashmore, 1984). All pLHCP derivatives carrying an exchanged amino acid X in position 222, termed [X222]pLHCP, have been described earlier (Paulsen and Kuttkat, 1993;Kuttkat et al, 1995). W222 in the present paper corresponds to W258 in Paulsen and Kuttkat (1993) and [X222]pLHCP in the present paper corresponds to W222X in Kuttkat et al (1995); (as for the numbering of amino acids see below).…”

Section: Methodsmentioning

confidence: 93%

“…This all-or-nothing effect has been interpreted as indicating a highly cooperative mode of complex siabilisation in which various parts of the protein and pigments undergo several mutual interactions, thus contributing to the overall stability (Cammarata and Schmidt, 1992;Paulsen and Hobe, 1992). In the C-terminal deletion series, the removal of up to ten amino acids does not significantly affect the stability of the pigment-protein complex ; only when the eleventh amino acid, W222, is removed, is the resulting protein unable to form ii monomeric complex with a stability comparable to that of native monomeric LHCII (Paulsen and Kuttkat, 1993; for the current model of LHCII and the numbering of amino acidx see Kuhlbrandt et al, 1994). However, when only W222 is exchanged in full-length LHCP, complex formation is not significantly affected, which suggests that W222 and (some of) the following ten amino acids exhibit an additive or mutually replaceable effect on complex stability.…”

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confidence: 99%

“…However, when only W222 is exchanged in full-length LHCP, complex formation is not significantly affected, which suggests that W222 and (some of) the following ten amino acids exhibit an additive or mutually replaceable effect on complex stability. Only when both components are removed does the protein lose its ability to stably bind pigments (Paulsen and Kuttkat, 1993).…”

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confidence: 99%

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The C‐Terminal Domain of Light‐Harvesting Chlorophyll‐a/b‐Binding Protein is Involved in the Stabilisation of Trimeric Light‐Harvesting Complex

Kuttkat

Hartmann

Hobe

et al. 1996

European Journal of Biochemistry

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Light‐harvesting chlorophyll a/b‐binding protein (LHCP) can be reconstituted with pigments in detergent solution to yield stable monomeric light‐harvesting chlorophyll a/b complex (LHCII). This reconstitution is not significantly affected when up to ten amino acids are deleted on the C‐terminus of LHCP or when a tryptophan, which is 11 positions from the C terminus (W222), is exchanged with other amino acids [Paulsen, H. & Kuttkat, A. (1993) Photochem. Photobiol. 57, 139–142]. Here we show that the exchange of W222 with histidine or glycine completely abolishes the ability of the protein to assemble into trimeric LHCII, either upon reconstitution of monomeric complexes in detergent solution or upon insertion into isolated thylakoids. It is concluded that part of the hydrophilic domain on the C‐terminus of LHCP, although not essential for the formation of stable monomeric LHCII, is involved in trimer formation. The different degree to which various amino acids in place of W222 affect trainer formation suggests that a hydrophobic amino acid is needed in this position.

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“…45), whereas digalactosyl diacylglycerol forms bilayers or reversed hexagonal phases depending on the acyl chain composition. Furthermore, modification of the C-terminal end of LHC II (46) has been shown to be important for stabilization of the protein, particularly of the pigment-protein complex.…”

Section: Discussionmentioning

confidence: 99%

Phosphorylation Controls the Three-dimensional Structure of Plant Light Harvesting Complex II

Nilsson¹,

Štys²,

Drakenberg³

et al. 1997

Journal of Biological Chemistry

100

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The most abundant chlorophyll-binding complex in plants is the intrinsic membrane protein light-harvesting complex II (LHC II). LHC II acts as a light-harvesting antenna and has an important role in the distribution of absorbed energy between the two photosystems of photosynthesis. We used spectroscopic techniques to study a synthetic peptide with identical sequence to the LHC IIb N terminus found in pea, with and without the phosphorylated Thr at the 5th amino acid residue, and to study both forms of the native full-length protein. Our results show that the N terminus of LHC II changes structure upon phosphorylation and that the structural change resembles that of rabbit glycogen phosphorylase, one of the few phosphoproteins where both phosphorylated and non-phosphorylated structures have been solved. Our results indicate that phosphorylation of membrane proteins may regulate their function through structural protein-protein interactions in surface-exposed domains. Light harvesting complex II (LHC II)1 is a major chlorophyllcontaining protein complex that accounts alone for half of the pigments involved in photosynthesis in plants. It is located mainly in appressed regions of the thylakoid membrane where it acts as the light-harvesting antenna for photosystem II (PS II). Reversible phosphorylation of LHC II is an established mechanism for redistribution of absorbed light energy between PS II and PS I. Phosphorylation of LHC II (giving LHC II(P)) is triggered by conditions where the plastoquinone pool of the photosynthetic electron transport chain becomes reduced (1). The kinase responsible for the phosphorylation of LHC II is not yet identified, although it is suggested that it is located in the core of photosystem II (2) or in contact with the cytochrome b 6 f complex (3, 4). LHC II(P) is found in the unappressed regions of the chloroplast thylakoid membrane and there acts as a lightharvesting antenna for photosystem I (PS I) (5-7). From electron crystallography of 2-dimensional crystals, a structure for the major part of non-phosphorylated LHC II has been described at 3.4-Å resolution (8). This structure reveals no information regarding the N-terminal domain that contains the phosphorylation site at position 5 (Thr); the protein backbone was traced only to residue 26 where it ends up close to the lipid membrane, consistent with the fact that the sequence between residues 21 and 29 (RVKYLGPF) (9) consists mainly of hydrophobic, aromatic, or charged amino acids. Aromatic residues are located at the membrane surface in structures of membrane proteins (10 -13), and residues Trp-16 and Tyr-17 of LHC II may also then form a point of contact with the membrane. LHC II has been shown to lose its ability to trimerize when more than the first 15 amino acids are removed from the apoprotein (14). At this site, specific lipid-protein interactions between the amino side chains and the lipid phosphatidylglycerol are involved in stabilization of the trimers (15), which implies that only the first 15 amino acid residues at the...

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PIGMENT COMPLEXES OF LIGHT‐HARVESTING CHLOROPHYLL a/b BINDING PROTEIN ARE STABILIZED BY A SEGMENT IN THE CARBOXYTERMINAL HYDROPHILIC DOMAIN OF THE PROTEIN*

Cited by 22 publications

References 12 publications

Trimerization and crystallization of reconstituted light-harvesting chlorophyll a/b complex.

Trimerization and crystallization of reconstituted light-harvesting chlorophyll a/b complex.

The C‐Terminal Domain of Light‐Harvesting Chlorophyll‐a/b‐Binding Protein is Involved in the Stabilisation of Trimeric Light‐Harvesting Complex

Phosphorylation Controls the Three-dimensional Structure of Plant Light Harvesting Complex II

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