Three-dimensional image reconstruction from ordered arrays of 70S ribosomes

Elucidating the medium-resolution structure of ribosomal particles: an interplay between electron cryo-microscopy and X-ray crystallography

“…The introduction of single-particle cryo-EM, in combination with powerful computational procedures such as angular reconstitution [3][4][5][6], has yielded detailed three-dimensional images of bacterial ribosomal particles and their functional complexes [7][8][9][10][11][12][13]. These three-dimensional reconstructions resemble the consensus topography observed by traditional negative-staining EM and confirm the existence of internal features, mainly associated with vacant spaces such as tunnels and hollows, detected about a decade ago in reconstructions from negatively stained crystalline arrays [14][15][16][17]. They have also been interpreted along similar lines, in conjunction with biochemical findings accumulated over three decades [18,19].…”

Approaching Atomic Resolution in Crystallography of Ribosomes

“…The small and large ribosomal subunits are well separated in most reconstructions of whole ribosomes, regardless of the source of the ribo some, the staining material, and the computational treatments (2,11,29,38,50). An empty space, comprising about 20% of the volume of the ribosome, located at the interphase between the two ribosomal subunits was clearly resolved in reconstructed images of 70S ribosomes from B. stearothermophilus (Figure 2) (50).…”

Crystallographic Studies on the Ribosome, a Large Macromolecular Assembly Exhibiting Severe Nonisomorphism, Extreme Beam Sensitivity and No Internal Symmetry

“…Initially, the models reconstructed from electron micrographs of tilt series of negatively stained crystalline arrays Arad et al, 1987) were exploited. Recently, these studies were extended to higher resolution, bene®ting from the impressive quantum jump in single-particle imaging by cryo electron microscopy (Stark et al, 1995;Frank et al, 1995), which yielded images resembling the common views observed by traditional electron microscopy and contain the features revealed in the lower-resolution reconstructed images from crystalline arrays.…”