Towards Ribosomal Structure at Peptide Level: Use of Crosslinking, Antipeptide Antibodies and Limited Proteolysis

The Structure of Ribosomal RNA: A Three‐Dimensional Jigsaw Puzzle

“…In the case of the ribosome, such data could subsequently be correlated with crystallographically derived structures of the proteins as they become available, and then fed back into the RNA models. A study of this nature would of course be an enormous and technically difficult undertaking, and it is not surprising that the number of amino acid contacts so far determined, either between pairs of proteins [120] or between individual proteins and the ribosomal RNA (eg. [121]), is still very small.…”

Preliminary NMR studies of Thermus thermophilus ribosomal protein S19 overproduced in Escherichia coli

“…There are two highly conserved regions in eubacterial and plant organelle sequences (33^12 and 66-78, Tth numbering) with His 68 conserved even in some eukaryotes. This amino acid residue was shown to be involved in crosslinking of pro- tein pair S13-S19 in both B. stearothermophilus and E. coli [9]. The molecular mass of the purified recombinant protein determined by SDS-PAGE [19] electrophoresis in denaturing conditions (10500) corresponds to that calculated from the amino acid sequence (10569).…”

“…Neutron diffraction data suggest that SI9 and S14 are very closely spaced [8]. Crosslinking data show that S13-S19 is one of the main protein pairs formed in the 30S ribosomal subunit from both Bacillus stearothermophilus and E. coli [9]. Here we report the cloning, sequencing and overexpression of the gene for rpS19 from T. thermophilus in E. coli as well as the purification of the recombinant protein and its preliminary NMR analysis.…”

Chloroplast ribosomes and protein synthesis.

“…The position of L23 in the E. coli ribosome has been controversial, with cross-linking and immunoelectron microscopy studies giving conflicting results (91,447). Kruft et al (328) propose that it has an elongated structure, with the N-terminal domain close to L29 at the base of the 50S subunit and the C-terminal domain on the ribosomal surface close to the peptidyltransferase center. Proteins equivalent to E. coli L23 are found in ribosomes from eubacteria, organelles, archaebacteria, and cytoplasmic ribosomes of eukaryotes (513,708).…”