Figure 6.

The accessible range of bn–b* complex stability constants containing mutant b* is extended by comparison with species of intermediate affinity. For example (lower right), the W38F complex was compared directly with the D35A and wild-type complexes, but the ΔΔG separating the latter complexes is so large that direct comparison is difficult. Comparison with multiple intermediates (upper left) improves the precision of measurement as required by the path independence of free energy. The numerical values are in kcal/mol.