Kinetics of inhibition of peptide bond formation on bacterial ribosomes.
A cell-free system derived from Escherichia coli has been used in order to study the kinetics of inhibition of peptide bond formation with the aid of the puromycin reaction in solution. A similar study has been carried out earlier on a solid support matrix with the same inhibitors. We find that the overall pattern of the kinetics of inhibition is the same in the two systems. At low concentrations of inhibitor there is a competitive phase of inhibition, whereas at higher concentrations of inhibitor the type of inhibition becomes mixed noncompetitive. The values of Ki of the competitive phase in the system in solution are: 5.8 microM (amicetin), 0.2 microM (blasticidin S), 0.5 microM (chloramphenicol), and 0.5 microM (tevenel). The inhibitors amicetin, blasticidin S, and tevenel interact with the ribosome in a reaction which is slower than that of the substrate puromycin, showing clear-cut characteristics of slow-onset inhibition in both systems. Chloramphenicol, on the other hand does not easily show such a delay in solution. It interacts with the ribosome relatively faster than the other three antibiotics. Despite this, chloramphenicol too shows characteristics of time-dependent inhibition.
Theocharis
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Laboratory of Biochemistry, School of Medicine, University of Patras, Greece.
Synetos
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Kalpaxis
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Drainas
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Coutsogeorgopoulos
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D001667:Binding, Competitive; D004926:Escherichia coli / Q000737:chemistry* / Q000187:drug effects; D007700:Kinetics; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology*; D011691:Puromycin / Q000494:pharmacology*; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000187:drug effects; D012270:Ribosomes / Q000737:chemistry / Q000187:drug effects*; D012996:Solutions
D010455:Peptides; D011500:Protein Synthesis Inhibitors; D012346:RNA, Transfer, Amino Acyl; D012996:Solutions; C066516:tRNA, N-acetylphenylalanyl-phenylalanine-; D011691:Puromycin