Enthalpy of hydrogen bond formation in a protein-ligand binding reaction.

Parallel measurements of the thermodynamics(free-energy, enthalpy, entropy and heat-capacity changes) of ligand binding toFK506 binding protein (FKBP-12) in H2O and D2O have been performed in an effortto probe the energetic contributions of single protein-ligand hydrogen bondsformed in the binding reactions. Changing tyrosine-82 to phenylalanine inFKBP-12 abolishes protein-ligand hydrogen bond interactions in the FKBP-12complexes with tacrolimus or rapamycin and leads to a large apparent enthalpicstabilization of binding in both H2O and D2O. High-resolution crystallographicanalysis reveals that two water molecules bound to the tyrosine-82 hydroxylgroup in unliganded FKBP-12 are displaced upon formation of the protein-ligandcomplexes. A thermodynamic analysis is presented that suggests that the removalof polar atoms from water contributes a highly unfavorable enthalpy change tothe formation of C=O...HO hydrogen bonds as they occur in the processes ofprotein folding and ligand binding. Despite the less favorable enthalpy change,the entropic advantage of displacing two water molecules upon binding leads to aslightly more favorable free-energy change of binding in the reactions withwild-type FKBP-12.