PMID:15461445
| Citation |
Seo, HS, Kiel, M, Pan, D, Raj, VS, Kaji, A and Cooperman, BS (2004) Kinetics and thermodynamics of RRF, EF-G, and thiostrepton interaction on the Escherichia coli ribosome. Biochemistry 43:12728-40 |
|---|---|
| Abstract |
Ribosome recycling factor (RRF) and elongation factor-G (EF-G) are jointly essential for recycling bacterial ribosomes following termination of protein synthesis. Here we present equilibrium and rapid kinetic measurements permitting formulation of a minimal kinetic scheme that accounts quantitatively for RRF and EF-G interaction on the Escherichia coli ribosome. RRF and EF-G (a) each form a binary complex on binding to a bare ribosome which undergoes isomerization to a more stable complex, (b) form mixed ternary complexes on the ribosome in which the affinity for each factor is considerably lower than its affinity for binding to a bare ribosome, and (c) each bind to two sites per ribosome, with EF-G having considerably higher second-site affinity than RRF. Addition of EF-G to the ribosome-RRF complex induces rapid RRF dissociation, at a rate compatible with the rate of ribosome recycling in vivo, but added RRF does not increase the lability of ribosome-bound EF-G. Added thiostrepton slows the initial binding of EF-G, and prevents both formation of the more stable EF-G complex and EF-G-induced RRF dissociation. These findings are relevant for the mechanism of post-termination complex disassembly. |
| Links |
PubMed Online version:10.1021/bi048927p |
| Keywords |
Coumarins; Escherichia coli/metabolism; Kinetics; Nucleotides/chemistry; Nucleotides/metabolism; Peptide Elongation Factor G/chemistry; Peptide Elongation Factor G/metabolism; Protein Binding/drug effects; Proteins/chemistry; Proteins/metabolism; Ribosomal Proteins; Ribosomes/chemistry; Ribosomes/metabolism; Spectrometry, Fluorescence; Thermodynamics; Thiostrepton/pharmacology |
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