CHEM 335 Biophysical Chemistry Lab report covering 3 experimental techniques that involved the Chemical Denaturation of Myoglobin or Hemoglobin Experiment 1 UV-visible spectroscopy Experiment 2 Fluorescence spectroscopy Experiment 3 Circular Dichroism Go through all documents 1 to 16. What you will find is that for each experimental technique – a prelab report, results, calculations and summary was done. The raw data is all provided. It is important that you read document 1 which breaks down how the report marking system will be. The goal of this lab report is to compare, discuss and explain what information was gathered by using the 3 techniques and in so doing answer the following questions in the discussion as mention in the document 1: Experiments 1-3 report • Does the concentration of guanadinium hydrochloride required to denature 50% of the proteins change from one technique to another? What do you expect and how does this compare to your data? • What aspects of denaturation does each technique study specifically? What does each of the techniques provide you information on, and how do the pieces of the story come together? • Does the data you have obtained using the three techniques agree? • Where are the positions of the tryptophan residues in your protein relative to the protein core, and how will this affect your denaturation curve? Discuss your results based on the tryptophan spectra under different conditions, as well as how myoglobin/hemoglobin differs from the proteins you studied? • How do the three ΔG values you have calculated for the fully-folded proteins compare? Is there a literature value that you can compare to? What does the ΔG at zero M guanadinium hydrochloride physically mean in relation to the stability of the protein you studied? • You performed a renaturation study. Does the protein completely refold? Discuss renaturation of myoglobin/hemoglobin, and proteins in general, based on results you obtained from each specific technique. • Some students are given myoglobin and others given hemoglobin to study. How does the protein you study differs from the one you did not study (myoglobin vs hemoglobin)? Since you only have half of the experimental results, you can theorize on what you know about the structure of the protein, and provide references to support your claim.
