Analysis of the puromycin binding site in the 70 S ribosome of Escherichia coli at the peptide level.
Photoinduced cross-linking of Escherichia coli 70 S ribosomes with [3H]puromycin has led to the labeling of ribosomal proteins S7, S14, S18, L18, and L29. Proteolytic fragmentation of these proteins and separation of the peptide mixtures by C18 reversed-phase high performance liquid chromatography resulted in six puromycin-labeled peptides which were applied to sequence analysis. The following peptides were found labeled: Pro1-Lys10 of S7, Ala28-Lys46 and Ala7-Lys11 of S14, Asp24-Lys29 of S18, Tyr64-Lys68 of L18, and Thr55-Lys60 of L29. For the first time the molecular environment of an antibiotic in the procaryotic ribosome is presented at the peptide level.
Max-Delbrück-Centrum für Molekulare Medizin, Berlin-Buch, Federal Republic of Germany.
O
O
Bischof
V
V
Kruft
B
B
Wittmann-Liebold
D000595:Amino Acid Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002851:Chromatography, High Pressure Liquid; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011691:Puromycin / Q000378:metabolism*; D012269:Ribosomal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*; D014316:Tritium
D010446:Peptide Fragments; D012269:Ribosomal Proteins; C029609:ribosomal protein L18; C035078:ribosomal protein L29; C039747:ribosomal protein S14; C041184:ribosomal protein S18; C037442:ribosomal protein S7; D014316:Tritium; D011691:Puromycin