Analysis of the reversible binding of virginiamycin M to ribosome and particle functions after removal of the antibiotic.
Type A synergimycins (VM) were shown to act catalytically and to induce two ribosomal alterations: (a) inability to promote polypeptide synthesis; (b) high-affinity binding of type B synergimycins (VS). A claim for irreversible binding of type A synergimycins to ribosomes has promoted the present reinvestigation. Submission of ribosomes from VM-treated bacteria to a purification procedure (supposed to remove the drug, according to a low association constant previously reported) yielded particles still holding residual VM. The formation of VM.ribosome complexes, more stable than previously inferred but without covalent linkage, was deduced from the extractability of complexed VM by organic solvents. Moreover, incubation of these complexes with increasing amounts of anti-VM immunoglobulins progressively restored ribosome activity in protein synthesis. Binding of VS to ribosomes, by fluorimetric titrations in the presence of substoichiometric concentrations of VM, was incompatible with catalytic action of type A synergimycins. Ribosomes from VM-treated bacteria displayed also a higher affinity for VS than did control ribosomes. This property did not disappear when ribosome.VM complexes were incubated with anti-VM IgG, nor when VM-IgG complexes were withdrawn from the reaction mixture by protein A-agarose binding. We can conclude that VM binding produces: (1) an inhibition of ribosome-promoted peptide bond formation, which occurs only in the presence of the drug; and (2) an increase of ribosome affinity for VS, which lasts after VM removal. The linkage of this drug with ribosomes is tight but reversible and its action is stoichiometric.
Nyssen
E
E
Microbiology & Genetics Unit, ICP, University of Lowvain Medical School, Brussels, Belgium.
Di Giambattista
M
M
Cocito
C
C
D000900:Anti-Bacterial Agents / Q000378:metabolism* / Q000494:pharmacology; D001426:Bacterial Proteins / Q000096:biosynthesis; D002458:Cell Fractionation; D004926:Escherichia coli / Q000378:metabolism / Q000648:ultrastructure*; D007074:Immunoglobulin G; D007158:Immunologic Techniques; D012270:Ribosomes / Q000187:drug effects / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014769:Virginiamycin / Q000008:administration & dosage / Q000378:metabolism* / Q000494:pharmacology
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D007074:Immunoglobulin G; D014769:Virginiamycin