Figure 3
From: Colorimetric Assay Reports on Acyl Carrier Protein Interactions
Analysis of the AcpP-TNB− reaction with FabF reveals the formation of AcpP-FabF complex and concomitant release of TNB2−. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) under non-reducing (A) and reducing conditions (B) indicate that the AcpP-FabF complex is selectively formed under non-reducing conditions. Ladder (lane 1), AcpP-TNB− (lane 2), FabF (lane 3), AcpP:FabF mixed in molar ratio of 0.2:1 (lane 4), 0.5:1 (lane 5), 1:1 (lane 6), 1.5:1 (lane 7), 2:1 (lane 8) and 3:1(lane 9). The release of TNB2− upon AcpP-FabF complex formation is observed by an increase in A412. (C) AcpP-TNB− (black); FabF (red); 0.2:1 (green); 0.5:1 (cyan); 1:1 (magenta); 1.5:1 (dark yellow); 2:1 (orange); and 3:1 (yellow). Inset shows the increase in A412 versus concentration of AcpP-TNB− added to FabF (average of two experiments).
