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Three-Dimensional Electron Microscopy of the Photosystem II Core Complex
Authors:Kouta Mayanagi  Takashi Ishikawa  Chikashi Toyoshima  Yorinao Inoue  Katsuyoshi Nakazato
Affiliation:aPhotosynthesis Research Laboratory, The Institute of Physical and Chemical Research (RIKEN), Wako, Saitama, 351-01, Japan;bInstitute of Molecular and Cellular Biosciences, University of Tokyo, Bunkyo-ku, Tokyo, 113, Japan;cPRESTO, Japan Science and Technology Corporation
Abstract:A three-dimensional image of the spinach photosystem II core complex composed of CP47, D1, D2, cytochromeb-559, andpsbI gene product was reconstructed at 20-Å resolution from the two-dimensional crystals negatively stained with phosphotungstate. Confirming the previous proposal, the crystal had ap22121symmetry. One PSII core complex was measured to be 80 × 80 Å in the membrane plane and 88 Å normal to it. The mass distribution was asymmetric about the lipid bilayer, consistent with predictions from the amino acid sequences. The lumenal mass consisted of three domains forming a characteristic triangular platform with another domain on top of it. Three stromal domains were smaller and linearly arranged. Due to strong stain exclusion in the hydrophobic core part of the lipid bilayer, the transmembrane region appeared to be imaged with a reversed contrast. Inverting the contrast resulted in a reasonable density distribution for that part. Thus, though the information on the transmembrane region is limited, the domain structure of the PSII core complex was revealed and allowed us to propose a model for the arrangement of subunits in the PSII core complex.
Keywords:electron microscopy   photosystem II   spinach   2-dimensional crystal   3-dimensional structure.
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