Three methods to assay inhibitors of ribosomal subunit assembly.
The inhibition of bacterial ribosomal subunit formation is a novel target for translational inhibitors. Inhibition of subunit biogenesis has been shown to be equivalent to the inhibition of protein biosynthesis for many antibiotics. This chapter describes three methods for examining the inhibition of subunit formation in growing bacterial cells. The first method permits the determination of the IC50 value for inhibition of assembly and protein synthesis. The second is a pulse and chase labeling procedure to measure the kinetics of subunit formation. The third procedure allows an examination of ribosome reformation after antibiotic removal as a part of the post-antibiotic effect. Together these procedures give a description of the relative inhibitory effects of an antibiotic on translation and subunit formation.
Champney
W Scott
WS
Department of Biochemistry and Molecular Biology, East Tennessee State University, Johnson City, TN, USA.
D004591:Electrophoresis, Polyacrylamide Gel; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D020128:Inhibitory Concentration 50; D008826:Microbial Sensitivity Tests / Q000379:methods; D014176:Protein Biosynthesis / Q000187:drug effects*; D011500:Protein Synthesis Inhibitors / Q000032:analysis* / Q000494:pharmacology; D054681:Ribosome Subunits, Large, Bacterial / Q000187:drug effects* / Q000378:metabolism*; D054680:Ribosome Subunits, Small, Bacterial / Q000187:drug effects* / Q000378:metabolism*; D013194:Staining and Labeling; D014529:Uridine / Q000378:metabolism
D011500:Protein Synthesis Inhibitors; D014529:Uridine