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An electron microscopic study of the archaeal feast/famine regulatory protein 5. Fourier filtration of images
Authors:Sanae A Ishijima  Lester Clowney  Masashi Suzuki
Institution:*)National Institute of Advanced Industrial Science and Technology (AIST), AIST Tsukuba Center 6-10, Higashi, 1-1-1, Tsukuba, Ibaraki 305-8566, Japan; **)Japan Science and Technology Agency (JST), Core Research for Evolutionary Science and Technology (CREST), Honmachi, 4-1-18, Kawaguchi Center Building, Kawaguchi, Saitama 332-0012, Japan
Abstract:Microcrystals of the feast/famine regulatory protein (FFRP) pot0434017 (FL11) were prepared by sonicating larger crystals. Using the microcrystals a cryo-electron micrograph was obtained, which showed a hexagonal packing of cylinder-like assemblies of FL11. This micrograph was processed by selecting, in the Fourier space, spots reflecting the crystal lattice, thereby removing the noise. The microcrystal was not totally free from distortion, and cylinders in local clusters adopted slightly different orientations. Thus, 25 hexagonal units closest to the ideal, each containing a cylinder at the center surrounded by six others, were manually selected. The averaged image was further processed to yield a perfect six-fold symmetry. These processed images, and some of the original images too, show bridges connecting cylinders, each corresponding to two pairs of N-domains, protruding from the two cylinders and contacting between them in the X-ray structure.
Keywords:Cryo-electron microscopy  Fourier analysis  hyper-thermophiles  image processing  protein electron microscopy
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