The pH dependence of stability of the activation helix and the catalytic site of GART.
We have predicted the free energy of unfolding for the pH-dependent helix-coil transition of the activation helix of GART using continuum electrostatic calculations and structural modeling. We have assigned the contributions of each element of secondary structure and of each ionizable residue, within and in the vicinity of the activation helix, to the stability of several fragments of GART that participate in the formation of the catalytic site. We demonstrate that the interaction of His121-His132 contributes 2.2 kcal/mol to the ionization free energy between pH 0 and approximately 6. We also show that the ionization state of a network of five histidines, His108, His119, His121, His132 and His137, and two aspartic acids Asp141 and Asp144, contributes approximately 12 kcal/mol to the stability of the catalytic site of GART, out of a total stability of 16 kcal/mol of the whole enzyme. These interactions are important for the formation of the catalytic site of GART.
D020134:Catalytic Domain; D004795:Enzyme Stability; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019877:Hydroxymethyl and Formyl Transferases / Q000737:chemistry*; D008958:Models, Molecular; D051026:Phosphoribosylglycinamide Formyltransferase; D017433:Protein Structure, Secondary; D013816:Thermodynamics
D019877:Hydroxymethyl and Formyl Transferases; D051026:Phosphoribosylglycinamide Formyltransferase