Haloarcula marismortui 50S subunit-complementarity of electron microscopy and X-Ray crystallographic information.

The large 50S subunit of the Haloarcula marismortui 70S ribosome was solved to 19 A using cryo-electron microscopy and single particle reconstruction techniques and to 9 A using X-ray crystallography. In the latter case, phases were determined by multiple isomorphous replacement and anomalous scattering from three heavy atom derivatives. The availability of X-ray and electron microscopy (EM) data has made it possible to compare the results of the two experimental methods. In the flexible regions of the 50S subunit, small differences in the mass distribution were detected. These differences can be attributed to the influence of packing in the crystal cell. The rotationally averaged power spectra of X-ray and EM were compared in an overlapping spatial frequency range from 60 to 13 A. The resulting ratio of X-ray to EM power ranges from 1 to 15, reflecting a progressively larger underestimation of the Fourier amplitudes by the electron microscope.

[1]  J. Frank,et al.  Three-dimensional reconstruction with contrast transfer function correction from energy-filtered cryoelectron micrographs: procedure and application to the 70S Escherichia coli ribosome. , 1997, Journal of structural biology.

[2]  W. O. Saxton,et al.  The correlation averaging of a regularly arranged bacterial cell envelope protein , 1982, Journal of microscopy.

[3]  Joachim Frank,et al.  A 9 Å Resolution X-Ray Crystallographic Map of the Large Ribosomal Subunit , 1998, Cell.

[4]  J. Frank,et al.  Conformational variability in Escherichia coli 70S ribosome as revealed by 3D cryo-electron microscopy. , 1999, The international journal of biochemistry & cell biology.

[5]  D H Bamford,et al.  Bacteriophage phi 6 envelope elucidated by chemical cross-linking, immunodetection, and cryoelectron microscopy. , 1992, Virology.

[6]  J. Frank,et al.  The ribosome at improved resolution: new techniques for merging and orientation refinement in 3D cryo-electron microscopy of biological particles. , 1994, Ultramicroscopy.

[7]  R P May,et al.  Inter‐protein distances within the large subunit from Escherichia coli ribosomes. , 1992, The EMBO journal.

[8]  J. Frank,et al.  A common-lines based method for determining orientations for N > 3 particle projections simultaneously. , 1996, Ultramicroscopy.

[9]  Joachim Frank,et al.  On the correction of the contrast transfer function in biological electron microscopy , 1995 .

[10]  J A Langer,et al.  A complete mapping of the proteins in the small ribosomal subunit of Escherichia coli. , 1987, Science.

[11]  J Frank,et al.  Escherichia coli 70 S ribosome at 15 A resolution by cryo-electron microscopy: localization of fMet-tRNAfMet and fitting of L1 protein. , 1998, Journal of molecular biology.

[12]  M van Heel,et al.  The 70S Escherichia coli ribosome at 23 A resolution: fitting the ribosomal RNA. , 1995, Structure.

[13]  H. Bartels,et al.  Characterization and preliminary attempts for derivatization of crystals of large ribosomal subunits from Haloarcula marismortui diffracting to 3 A resolution. , 1991, Journal of molecular biology.

[14]  R. H. Wade.  A brief look at imaging and contrast transfer , 1992 .

[15]  R. Wade,et al.  Electron microscope transfer functions for partially coherent axial illumination and chromatic defocus spread , 1977 .

[16]  B. Wittmann-Liebold.  Ribosomal Proteins: Their Structure and Evolution , 1986 .

[17]  G. Kramer,et al.  Structure, Function, and Genetics of Ribosomes , 1986, Springer Series in Molecular Biology.

[18]  J. Frank,et al.  A model of protein synthesis based on cryo-electron microscopy of the E. coli ribosome , 1995, Nature.