19F nuclear magnetic resonance spectroscopy study of the complex of 2-benzyl-4-oxo-5,5,5-trifluoropentanoic acid and carboxypeptidase A.

(dl)-2-Benzyl-4-oxo-5,5,5-trifluoropentanoic acid is a strong transition state analog inhibitor of the zinc protease carboxypeptidase A. 19F NMR spectroscopy of the aqueous solution of this inhibitor shows the hydrate of the ketone carbonyl to be the major species, with a shift of -9.95 ppm. As the pH is varied from 4.9 to 13.1, a 1.53 ppm downfield shift occurs, giving a pK alpha of 11.10. When excess inhibitor is added to the enzyme, a new, bound peak appears at -8.84 ppm, in addition to the free hydrate peak. Spectra taken at pH's from 4.90 to 9.15 show no change in the position of the bound resonance; from 9.15 to 12.15, a 0.26 ppm upfield shift occurs. The interpretation is that the monoanion of the hydrate is the form that binds to the enzyme.