Trimerization and crystallization of reconstituted light-harvesting chlorophyll a/b complex.
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.
Hobe
S
S
Botanisches Institut III der Universität, Munich, Germany.
Prytulla
S
S
Kühlbrandt
W
W
Paulsen
H
H
D000595:Amino Acid Sequence; D003460:Crystallization; D018360:Crystallography, X-Ray; D045342:Light-Harvesting Protein Complexes; D008055:Lipids; D008969:Molecular Sequence Data; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000648:ultrastructure; D045332:Photosystem II Protein Complex; D011487:Protein Conformation
D045342:Light-Harvesting Protein Complexes; D008055:Lipids; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex