1. USE A COMPUTER TO SOLVE THIS PROBLEM. A common strategy for studying enzymatic mechanisms is to measure the pH-profile [log(kcm) versus pH] to identify catalytically essential ionizable amino acids in the active site of an enzyme. Observed pKA-values in this analysis can be tentatively assigned based on the measured pKa of the free amino acid in solution (shown in Table 1). However, a significant problem in such assignments is that the local environment of the ionizable group within the enzymatic site can vastly shift the pka-value by up to 5-pH units (5-orders of magnitude). For instance, the typical pKA for an Asp residue is -3.5, yet Asp102 of Chymotrypsin participates in a charge-relay hydrogen bonding network with His75 and Ser195 within the active site of Chymotrypsin. Thus, the measured pKa-value for Asp102 within this catalytic triad is increased to 7.0. Table 1. Ionizable groups present potentially present in an enzyme. Amino Acid Glu/Asp Lys Group a-carboxyl (C-terminus) By-carboxyl (R-group) 6.28 +6.28 28.9-314 41.9-54.4 41.9-54.4 272-29.3 3.0-5.0 5.5-7.0 7.5-8.5 a-amino (N-terminus) sulfhydryl phenolic-(OH) 9.5-10.6 8.0-8.5 98-10.5 Tyr 25.1 Aguanidinium11.6-126 502-54.4 Derive an expression to relate the observed pKA value at a specific temperature to the enthalpy and entropy of ionization (AH and AS). b. The pH-profile of an unknown enzyme exhibits (2) ionizable groups in [loghon)-pH] profiles. Use the expression derived afove to analyze the temperature dependence shown in Table 2 and assign each ionization to a specific residue based on pKa-value, Aloe and AS c. What chemical arguments can be made regarding the values of pKa-value, AHe and AS Table 2. Temperature dependence of enzymatic ionizable groups pparent pk 5.50 to apparentpKu 10 15 20 25 30 35 6.51 6.44 6.37 6.30 6.23 6.17 6.11 10 5.54 5.55 5.56 5.57 30 35 MacBook A