Figure 3.
Influence of changing monovalent ion concentration and temperature on the 50S subunit's pH-dependent DMS reactivity at A2451 and ability to perform peptidyl transfer. Unm, unmodified ribosomes. (A) Heat activation of 50S subunits in K+ (lanes 6–8) or NH
(lanes 12–14) results in loss of pH-dependent DMS reactivity at A2451. (B) Heat incubation in the absence of monovalent cation (lanes 6–8) or in the presence of Na+ (lanes 12–14) does not result in loss of pH-dependent DMS reactivity at A2451. (C) Increasing concentrations of K+ (lanes 3–5) or NH
(lanes 6–8) at constant pH (6.5) at 0°C results in increasing DMS reactivity of A2451. (D) 50S subunits shown to exhibit pH-dependent DMS reactivity at A2451 cannot perform peptidyl transferase as assayed by the fragment reaction. 50S preincubation conditions are as shown.
