Ribosome structure: binding site of macrolides studied by photoaffinity labeling.
The macrolide antibiotics carbomycin A, niddamycin, and tylosin have been radioactively labeled by reducing their aldehyde group at the C-18 position. Dihydro derivatives with specific activities around 2.5 Ci/mmol can be obtained that, although partially affected in their activity, still bind to the ribosomes with high affinity. The presence in the chemical structure of these antibiotics of alpha-beta-unsaturated ketone groups makes them photochemically reactive, and by irradiation above 300 nm, covalent incorporation of the radioactive dihydro derivatives into ribosomes has been achieved. The covalent binding seems to take place at the specific binding sites for macrolides as deduced from binding saturation studies and competition experiments with unmodified drugs. Analysis of the ribosomal components labeled by the drugs indicated that most radioactivity is associated with the proteins L27, L2, and L28 when 50S subunits are labeled, and with L27, L2, L32/33, S9, and S12 in the case of 70S ribosomes. These results agree well with a model of macrolides' mode of action that assumes an interaction of the drug at the peptidyl transferase P site that would block the exit channel for the growing peptide chain.
D000345:Affinity Labels; D000900:Anti-Bacterial Agents / Q000378:metabolism*; D001665:Binding Sites; D004926:Escherichia coli / Q000648:ultrastructure; D007783:Lactones; D010777:Photochemistry; D012269:Ribosomal Proteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism / Q000648:ultrastructure*
D000345:Affinity Labels; D000900:Anti-Bacterial Agents; D007783:Lactones; D012269:Ribosomal Proteins