For many years methods to describe the structure of the transition state for a reaction have been sought. Most commonly these structures have been inferred from kinetic isotope effects. We report here for the first time the application of transition state pK(a) values to describe the relationship between molecular recognition and the transition state for the catalytic mechanism of papain. The background to the theory is presented and applied to the reactions of papain with a series of trimethylene disulphide reactivity probes. The common feature of these reactions is a loss in reactivity on ionization of the imidazolium cation for those probes containing molecular recognition features and an increase in reactivity on ionization of the electrostatic switch residue. The use of transition state pK(a) values enhances this information by providing details regarding the protonic distribution within the transition state. This has led to the reconsideration of the effect of the electrostatic switch ionization and the role of the hydrogen bond formed between the catalytic-site imidazolium cation and the leaving group of the reaction in the catalytic mechanism of papain.