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ST. EDWARDS CHEM 4343 - Quiz

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CChheemm 44334433 Quiz Nov 2008 Name : Answer each of the following five parts for a total of 100 points. 1 (i) Complete the enzymatic mechanism for each of the following reaction steps: (ii) the chemical drawing on the right shows the active site of an enzyme, with the three amino acid residues in the active site labeled as AA1, AA2 and AA3. The enzyme is bound to a tripeptide (Lys-Gly-Gly) substrate.Complete as many steps of the catalytic mechanism as possible andof this reaction. determine, giving your reasoning, the pH profile (iii) two different substrates for a given enzymatic reaction are shown below, along with the kinetic data for each reaction. Determine, giving your reasoning, which substrate would produce the most product in a given period of time ( if necessary you can assume that the substrate concentration was 0.01 mM. AA1 AA2 AA3 H2O(iv) In the hydrolysis reaction shown below the same substrate is shown bound to three different enzymes, the normal or wild type enzyme (wt) and two mutants that have one amino acid changed in the active site. Determine, giving your reasoning, which column of kinetic data belongs to which mutant. [S] V(wt) V V (mM) ([S]/s) ([S]/s) ([S]/s) 0.5 33 11 17 1 50 20 25 2 67 33 33 10 91 71 45 20 95 83 48 50 98 93 49 75 99 95 49 (v) In the plot shown on the right the enzyme kinetic data was obtained for the normal enzyme reacting with the normal substrate in the absence of inhibitor, i.e. the drug. Determine a value for VMAX and KM. On the diagram that is given draw the plot for the reaction of a) mutant enzyme + normal substrate b) normal enzyme + substrate+ Drug normal enzyme active site mutant enzyme active siteBonus (10 pts) : For the The drawing to the right shows a substrate interacting with Val82 from the enzyme HIV protease. Two inhibitors, drug A and drug B were developed to treat the disease. For the kinetic plot shown below: a) identify, giving your reasoning, which line belongs to the which assay i.e. enzyme+substrate, enzyme+substrate+Drug A, enzyme+substrate+Drug B b) If you are told that a mutant enzyme reverses the effectiveness of these drugs suggest, giving your reasoning, a plausible amino acid difference in this mutant


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