25. a. What molecular features would make a molecule hydrophilic? (2 points)

b. What molecular features would make a molecule amphipathic? (3 points)

26. a. If you had a protein and wanted to make it more positively charged, what two amino acids could you add to its sequence? (2 points)

b. What would you replace them with if you wanted to make it more negative? (2 points)

27. How would you define "Domains" in protein? (3 points)

28.  List three different kinds of domains in protein. (3 points)

29. Why does NaCl dissolve in water? (3 points)

30. Why does water expand when it freezes? (3 points)

Question 31.

Assume that you are trying to analyze what you believe to be a pure protein. You carry out the following procedures:

A. When you elute from an ion exchange column, you get a single peak of material. On what basis is this separation occurring? (1 point)

B. When this material is run on Sephadex, it elutes as two peaks, one with an apparent molecular weight of 80 kD, and the other with an apparent molecular weight of 40 kD. On what basis is this separation occurring? (1 point)

C. If you electrophorese the protein in the presence of SDS, it migrates as two bands with apparent molecular weights of 40 and 80 kD. On what basis is this separation occurring? (1 point)

To your surprise, if you treat the sample with β-mercaptoethanol and then electrophorese in SDS, you now get a single band migrating with an apparent molecular weight of 40 kD. 

D. What was in your original sample? Explain your reasoning. Remember, you need to account for all of the previous results. (2 points)

Question 32. Fill in the Following Table:

NB: "Gravity" is NOT the answer to any of these questions!

 

 Separation Technique, Force that moves the sample (1 point)

Property of the sample molecules by which they are separated (2 points)

What provides resistance to movement? (2 points)

 

Of

1. Ultracentrifuge -

   2. Electrophoresis in Acrylamide Gel -

(no SDS)

3. Electrophoresis in Acrylamide Gel with SDS -

4. Gel Filtration Chromatography -

5. Ion Exchange Chromatography -

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