Figure 3.
2D 19F-19F NOESY exchange spectra of 19F-Man2/Man3 in the presence of [CVNΔA]ssm and CVNmDB protein. A) 19F-Man2 (200μM) bound to [CVNΔA]ssm(left panel; tmix=0.1 s), and the exchange curve of bound/bound over free/bound intensity ratio (IBB/IBF) versus mixing time (right panel) tmix: 0.05, 0.1, 0.3, 0.5 s. B) 19F-Man2 bound to CVNmDB (left panel; tmix=0.05 s) and the exchange curve of bound/bound over free/bound intensity ratio (IBB/IBF) versus mixing time (right panel) tmix: 0.025, 0.05, 0.1, 0.2 s. C) 19F-Man3 (50 μM) bound to [CVNΔA]ssm (left panel; tmix=0.1 s) and the exchange curve of bound/bound over free/bound intensity ratio (IBB/IBF) versus mixing time (right panel) tmix: 0.1, 0.3, 0.5, 0.7, 0.9 s. D) 19F-Man3 (50 μM) bound to CVNmDB (left panel; tmix=0.05 s) and the exchange curve of bound/bound over free/bound intensity ratio (IBB/IBF) versus mixing time (right panel) tmix: 0.05, 0.1, 0.15, 0.3 s. All spectra were recorded at 280 K for 1:1 molar ratios. The auto-correlation (diagonal) peaks IFF, and IBB arise from the free and bound states, while cross-correlation (off-diagonal) peaks IFB=IBF report on the exchange between free and bound states.