Reports must be submitted to Canvas by the start of your next lab.

Refer to the general guidelines for Mini Lab Reports on Canvas, found here (link opens in a new tab): Mini Lab Report Guidelines


Results Section

For each of the three mutations you designed during lab, address the following:

  • State the residue that you changed and what amino acid you changed it to (e.g. V34K). Include a figure showing the change in the structure. Please make sure that the PyMOL figures you use in the lab report have a white background – this will make them significantly easier to evaluate. Remember that figures need to have informative titles and legends. Your legend should clearly state what is shown in each color and what is depicted as sticks/spheres/etc.
  • Justify why you chose to change that amino acid and why you selected the mutant residue that you did. Use your knowledge of the structure from the PyMOL session in Lab 1 and the biochemical properties of the amino acids (Fig. 1.7 or any general biochemistry textbook) to inform your answer. (It is ok to mention the expected phenotype here, if necessary, but that should mainly be discussed in the Discussion Section of the report.)
  • Comment on clashes that appear with your mutated residues.
  • Study the effect of the mutations you picked on the ligand-binding structure. (The ligand is dansylamide, DNSA). Do they seem compatible, or do the mutations create clashes?


Discussion Section

What effect on HCAII function do you expect to observe for each of the three mutations? Consider the following:

  • Different functionality of the new residue (is it a conservative or non-conservative mutation?)
  • Effects on structure
  • Effects on enzymatic activity
  • Effects on ligand-binding

What are the next steps? How would you test the function of the mutants?


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Biochemistry 551 (Online Version) Lab Manual Copyright © by Lynne PROST is licensed under a Creative Commons Attribution 4.0 International License, except where otherwise noted.

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