ON THE PH-DEPENDENCE OF AMIDE PROTON-EXCHANGE RATES IN PROTEINS

We have analyzed the pH dependencies of published amide proton exchange rates (k(ex)) in three proteins: bovine pancreatic trypsin inhibitor (BPTI), bull seminal plasma proteinase inhibitor IIA (BUSI IIA), and calbindin D-9K. The base-catalyzed exchange rate constants (k(OH)) of solvent exposed amides in BPTI are lower for residues with low peptide carbonyl exposure, showing that the environment around the carbonyl oxygen influences k(OH). We also examined the possible importance of an exchange mechanism that involves formations of imidic acid intermediates along chains of hydrogen-bonded peptides in the three proteins. By invoking this ''relayed imidic acid exchange mechanism,'' which should be essentially acid-catalyzed, we can explain the surprisingly high pH(min) (the pH value at which k(ex) reaches a minimum) found for the non-hydrogen-bonded amide protons in the beta-sheet in BPTI. The successive increase of pH(min) along a chain of hydrogen-bonded peptides from the free amide to the free carbonyl, observed in BPTI, can be explained as an increasing contribution of the proposed mechanism in this direction of the chain. For BUSI IIA (pH 4-5) and calbindin D-9K (pH 6-7) the majority of amide protons with negative pH dependence of k(ex) are located in chains of hydrogen-bonded peptides; this situation is shown to be consistent with the proposed mechanism.