Study of mammalian ribosomal protein reactivity in situ. I. - Effect of 2-methoxy-5-nitrotropone on 40S and 60S subunits.
Liver ribosomes and subunits were reacted with increasing concentrations of 2-methoxy-5-nitrotropone. At low reagent concentrations (0.3 mM), the molar uptake by 60S subunits was more efficient than the uptake by 40S subunits, and the amount of reagent bound to 80S ribosomes was less than that bound to both free subunits considered together. At higher reagent concentrations, the molar uptake of both subunits was equivalent. Subunits and ribosomes remained fully active when reacted with up to 0.3 mM and 1 mM of the reagent, respectively. With 2 mM of the reagent, both subunits were half inactivated, although their sedimentation characteristics were unaltered. The reactivity of each ribosomal protein was assessed by two-dimensional gel electrophoresis and quantitative measurement of the unmodified proteins. From these results, considered together with the uptake characteristics and the inactivation curves, a number of tentative conclusions about ribosome topography can be drawn. The over-all sensitivity of the 60S subunits to the reagent is higher than that of the 40S subunits. Both subunits undergo a conformational change when they combine to form 80S ribosomes. Proteins S18, S20, S28 and L5, L9, L11, L15, L16, L25, L29, L30, L31, L34, L37 have NH2 groups exposed in native subunits. These groups are not essential for subunit function.
D000818:Animals; D002474:Cell-Free System; D003864:Depression, Chemical; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D007202:Indicators and Reagents; D008099:Liver; D008297:Male; D009690:Nucleic Acid Conformation; D014176:Protein Biosynthesis / Q000187:drug effects; D011487:Protein Conformation; D051381:Rats; D012269:Ribosomal Proteins / Q000302:isolation & purification; D012270:Ribosomes / Q000187:drug effects*; D013329:Structure-Activity Relationship; D014326:Tropanes / Q000494:pharmacology
D007202:Indicators and Reagents; D012269:Ribosomal Proteins; D014326:Tropanes; C008402:2-methoxy-5-nitrotropone