Lab 11: FINAL LAB REPORT

FINAL LAB REPORT

There is no Mini Lab Report for Lab 11. What follows are some suggestions for what to include from this lab in your final report.

Methods

You again need to write a detailed methods section. Include:

• Final concentration of HCAII, DNSA, AZ
• Equations for calculating for K_D for both DNSA and AZ and make sure to define the values for F_max and F_min.

Results

• A graph of the fraction saturation (r) vs [DNSA]_free, and graph of r vs [AZ]_free
• Describe the results of your plots in the text including relevant values
• Be sure to include measurements of error and p-values, where appropriate

Discussion

• For which ligand, Dansyl amide or acetazolamide, does HCAII have a tighter affinity? By how much? Based upon the structure of DNSA and AZ, which would you expect to have higher affinity and why?
• How do your K_D values compare to that of the literature? What are some reasons why your values could differ from the published results?
  • Here is one useful reference on this topic (pdf opens in a new tab): Structural Basis of Inhibitor Affinity to Variants of Human Carbonic Anhydrase II
• How does the HCAII mutation you studied affect ligand binding? Discuss the relative affinity of DNSA and AZ to wt HCAII and the mutant. Your answer should include information about the chemistry of the amino acid in the wild type protein and the mutation you made as well as a statement about how the mutation affected the structure of the protein and thus its ability to bind the ligands. Use your results to support your conclusions.