Two-dimensional crystallization and cryo-electron microscopy of photosystem II

Journal of Molecular Biology
K NakazatoY Inoue

Abstract

Two dimensional (2D) crystals of photosystem II (PSII) were obtained from n-heptyl-beta-D-thioglucoside-solubilized monomers of spinach PSII complex by conventional detergent dialysis. The 2D crystals were either large cylindrical vesicles (1 to 2 micrometer by 4 to 6 micrometer as flattened vesicles) or large monolayer sheets (ca. 1 micrometer X 1 micrometer), both suitable for cryo-electron microscopy. Images of unstained crystals embedded in ice were recorded using low-dose microscopy and analyzed by digital image processing. Both types of crystals had the same unit cell size and the same packing arrangement of PSII particles. The plane group was p22(1)2(1) and the unit cell was rectangular with dimensions of 16.7 nm X 15.3 nm containing four monomers (two face-up and two face-down). SDS-PAGE and immunoblot analyses of the 2D crystal indicated that the constituent subunits of particles in the 2D were CP47, D1, D2, cytochrome b-559 and psbI protein. A projection map of 20 A resolution revealed that each monomer has asymmetrical shape with a length of 8.1 nm and a maximal width of 7.5 nm consisting of four areas of density. Two high-density areas with similar sizes were located close to each other to form a roughly rectangular...Continue Reading

Citations

Jun 1, 1997·Annual Review of Plant Physiology and Plant Molecular Biology·Ben HankamerEgbert J. Boekema
May 8, 2001·Annual Review of Biophysics and Biomolecular Structure·K H Rhee
May 12, 2007·Photosynthesis Research·Jan KernAthina Zouni
Jul 20, 2007·Photosynthesis Research·Jan Kern, Gernot Renger
Nov 1, 1996·Photosynthesis Research·G TsiotisD Ghanotakis
Jun 10, 1998·Journal of Structural Biology·T Walz, N Grigorieff
Aug 1, 1997·Current Opinion in Structural Biology·J B HeymannA Engel
Feb 13, 2001·Micron : the International Research and Review Journal for Microscopy·G Mosser
Nov 25, 2000·Biochimica Et Biophysica Acta·J RigaudJ Ranck
Jun 19, 1998·Biochimica Et Biophysica Acta·M K Lyon

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