Figure 1.

Synergistic degradation of S. pneumoniae R6 cell walls by Cpl-711 and PL3. (A) 1.3μM solution (≈50 μg/ml) of either PL3, Cpl-711, a mixture of both enzymes (MIX, either 1:10 or 10:1, respectively 0.13μM PL3 + 1.3μM Cpl-711, and 1.3μM PL3 + 0.13μM Cpl-711), or just PB as a no-treatment control and spectrophotometric blank read, were added onto pelleted RBB-stained R6 cell walls adjusting to A₅₉₅ ≈ 0.5. Degradation products were measured spectrophotometrically after sample centrifugation at indicated time points. (B) RBB-stained cell walls were pre-treated with 1.3μM PL3 (Left panel) or Cpl-711 (Right panel) in PB for 1 h, and reactions were stopped by heat inactivation (80°C, 15 min). Incubation was then resumed at 37°C adding either 1.3μM PL3 (black circles), 1.3μM Cpl-711 (white squares) or an equivalent PB volume (gray diamonds) at the time indicated by the arrows.