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Crystallization of Escherichia coli ribosomes
Author(s) -
Wittmann H.G.,
Müssig J.,
Piefke J.,
Gewitz H.S.,
Rheinberger H.R.,
Yonath A.
Publication year - 1982
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(82)80739-4
Subject(s) - physics , planck , astrophysics
The knowledge of the structure of the ribosome is an essential requirement to reveal its role at the molecular level in the process of protein biosynthesis. This information is being obtained by a battery of chemical, physical, immunological and genetic methods (for reviews see [1]). Important methods for the study of the three-dimensional structure of the ribosomes are X-ray crystallography and image reconstruction. Two-dimensional ordered sheets of ribosomes from a few eukaryotic species are found under special conditions in vivo [2-4] or in cell homogenates [5]. Small two-dimensional arrays of E. coli ribosomal subunits were obtained in vitro [6,7]. The first three-dimensional crystals of ribosomal particles were obtained in vitro from the large subunit of the Bacillus stearothermophilus ribosome [8,9]. These have been characterized by three-dimensional image reconstruction studies [10]. Here we report the in vitro crystallization of E. coli ribosomes. We have obtained ordered threedimensional microcrystals. After washing and redissolving, the crystalline ribosomes sediment with a coefficient of 70 S in a sucrose gradient and are biologically active in the poly(U) in vitro system. Electron micrographs of thin sections of the crystals show regular arrangement of the ribosomes, and their optical diffraction patterns extend out to about 6.5 nm. Thus three-dimensional image reconstruction studies can be carried out. 2. MATERIALS AND METHODS