Dynamics of trimethoprim bound to dihydrofolate reductase.

Abstract
The conformation of a small molecule in its binding site on a protein is a major factor in the specificity of the interaction between them. In this paper, we report the use of 1H and 13C NMR spectroscopy to study the fluctuations in conformation of the anti-bacterial drug trimethoprim when it is bound to its "target", dihydrofolate reductase. 13C relaxation measurements reveal dihedral angle changes of .+-. 25.degree. to .+-. 35.degree. on the subnanosecond time scale, while 13C line-shape analysis demonstrates dihedral angle changes of at least .+-. 65.degree. on the millisecond time scale. 1H NMR shows that a specific hydrogen bond between the inhibitor and enzyme, which is believed to make an important contribution to binding, makes and breaks rapidly at room temperature.