Cross-linking of streptomycin to the 50S subunit of Escherichia coli with phenyldiglyoxal.
[3H]Dihydrostreptomycin was covalently linked to the 50S subunit of Escherichia coli K12A19 with the bifunctional cross-linking reagent phenyldiglyoxal. The cross-linking was abolished under conditions that prevent the specific interaction of streptomycin with the ribosome. The binding primarily involved the ribosomal RNA and also a limited number of proteins, namely, L2, L6, and L17. This suggests that the binding domain for streptomycin is close to the peptidyl transferase center, in the valley between the central protuberance and the wider lateral protuberance of the 50S subunit. This domain faces the binding domain for streptomycin which we have previously characterized on the 30S subunit [Melançon, P., Boileau, G., & Brakier-Gingras, L. (1984) Biochemistry 23, 6697-6703]. Our results indicate that the 50S subunit is involved in the binding of streptomycin to the bacterial ribosome, in addition to the 30S subunit which is generally considered as the specific target of the antibiotic. They are consistent with the occurrence of a single binding site for streptomycin on the ribosome, comprised of regions of both subunits.
D000447:Aldehydes / Q000494:pharmacology*; D002458:Cell Fractionation; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000378:metabolism*; D008970:Molecular Weight; D010658:Phenylglyoxal / Q000031:analogs & derivatives / Q000494:pharmacology*; D012269:Ribosomal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D013307:Streptomycin / Q000378:metabolism*
D000447:Aldehydes; D003432:Cross-Linking Reagents; D012269:Ribosomal Proteins; C018360:1,4-phenyldiglyoxal; D010658:Phenylglyoxal; D013307:Streptomycin