The three-dimensional structure of a photosystem II core complex determined by electron crystallography.
BACKGROUND
Photosystem II (PSII) is a multisubunit protein complex which is embedded in the photosynthetic membranes of plants. It uses light energy to split water into molecular oxygen and reducing equivalents. PSII can be isolated with varying degrees of complexity in terms of its subunit composition and activity. To date, no three-dimensional (3-D) structure of the PSII complex has been determined which allows location of the proteins within the PSII complex and their orientation in relation to the thylakoid membrane.


RESULTS
Two-dimensional (2-D) PSII core complex crystals composed of the two reaction centre proteins, D1 and D2, two chlorophyll-binding proteins, CP47 and CP43, cytb559 and associated low molecular weight proteins were formed after reconstituting the isolated complex into purified thylakoid lipids. Electron micrographs of negatively stained crystals were used for 2-D and 3-D image analyses. In the resulting maps, the PSII complex is composed of two halves related by twofold rotational symmetry, thus, confirming the dimeric nature of the complex; each monomer appears to contain five domains. Comparison of the 3-D images with platinum shadowed images of the crystals allowed the likely lumenal and stromal surfaces of the complex to be identified and regions contained within the membrane to be inferred. The projection structure of 2-D crystals of a smaller CP47-D1-D2-cytb559 complex was used to identify the domains apparently associated with CP43.


CONCLUSIONS
The results indicate that PSII probably exists as a dimer in vivo. The extensive proteinaceous protrusions from the lumenal surface have been tentatively assigned to hydrophilic loops of CP47 and CP43; the positioning of these loops possibly implies their involvement in the water-splitting process.
Morris
E P
EP
Wolfson Laboratories, Department of Biochemistry, Imperial College of Science, Technology & Medicine, London, UK.
Hankamer
B
B
Zheleva
D
D
Friso
G
G
Barber
J
J
D003460:Crystallization; D019281:Dimerization; D007091:Image Processing, Computer-Assisted; D045342:Light-Harvesting Protein Complexes; D008051:Lipid Bilayers; D008854:Microscopy, Electron / Q000379:methods*; D008958:Models, Molecular; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry* / Q000378:metabolism; D045332:Photosystem II Protein Complex
D045342:Light-Harvesting Protein Complexes; D008051:Lipid Bilayers; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; C070970:photosystem II, chlorophyll-binding protein, CP-47