A congeneric series of 1,2-azaborine ligands was used to study the relationship between the steric demand of the ligand and hydrogen bonding strength in the context of ligand-protein binding using engineered T4 lysozymes as the model biomacromolecules. The hydrogen bonding strength values were extracted from experimentally accessible binding free energies using the Double Mutant Cycle analysis. With the increasing steric demand, the NH…102Q hydrogen bonding interaction is weakened; however, this weakening of the hydrogen bonding interaction occurs in discrete steps rather than in an incremental fashion.