Specific binding of ribosome recycling factor (RRF) with the Escherichia coli ribosomes by BIACORE.
The direct assays on Biacore with immobilised RRF and purified L11 from E. coli in the flow trough have shown unspecific binding between the both proteins. The interaction of RRF with GTPase domain of E. coli ribosomes, a functionally active complex of L11 with 23S r RNA and L10.(L7/L12)4 was studied by Biacore. In the experiments of binding of RRF with 30S, 50S and 70S ribosomes from E. coli were used the antibiotics thiostrepton, tetracycline and neomycin and factors, influencing the 70S dissociation Mg2+, NH4Cl, EDTA. The binding is strongly dependent from the concentrations of RRF, Mg2+, NH4Cl, EDTA and is inhibited by thiostrepton. The effect is most specific for 50S subunits and indicates that the GTPase centre can be considered as a possible site of interaction of RRF with the ribosome. We can consider an electrostatic character of the interactions with most probable candidate 16S and 23S r RNA at the interface of 30S and 50S ribosomal subunits.
Todorova
Roumiana T
RT
Institute of Biophysics, Bulgarian Academy of Sciences, 1113 Sofia, Bulgaria. todorova@obzor.bio21.bas.bg
Saihara
Yukari
Y
D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D015374:Biosensing Techniques; D004926:Escherichia coli / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D009355:Neomycin / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism; D012269:Ribosomal Proteins; D012270:Ribosomes / Q000378:metabolism*; D013752:Tetracycline / Q000494:pharmacology; D013883:Thiostrepton / Q000494:pharmacology; D013997:Time Factors
D011506:Proteins; D012269:Ribosomal Proteins; C087267:ribosome releasing factor; D012343:RNA, Transfer; D013752:Tetracycline; D013883:Thiostrepton; D009355:Neomycin; D008274:Magnesium