ABSTRACT
Cytosolic serine hydroxymethyltransferase has been shown previously to exhibit both broad substrate and reaction specificity. In addition to cleaving many different 3-hydroxyamino acids to glycine and an aldehyde, the enzyme also catalyzes with several amino acid substrate analogs decarboxylation, transamination, and racemization reactions. To elucidate the relationship of the structure of the substrate to reaction specificity, the interaction of both amino acid and folate substrates and substrate analogs with the enzyme has been studied by three different methods. These methods include investigating the effects of substrates and substrate analogs on the thermal denaturation properties of the enzyme by differential scanning calorimetry, determining the rate of peptide hydrogen exchange with solvent protons, and measuring the optical activity of the active site pyridoxal phosphate. All three methods suggest that the enzyme exists as an equilibrium between "open" and "closed" forms. Amino acid substrates enter and leave the active site in the open form, but catalysis occurs in the closed form. The data suggest that the amino acid analogs that undergo alternate reactions, such as racemization and transamination, bind only to the open form of the enzyme and that the alternate reactions occur in the open form. Therefore, one role for forming the closed form of the enzyme is to block side reactions and confer reaction specificity.
