https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3504993/
  1. Your assigned mutant for this question is VA from the Richards et al. article. Answer the following

a. What actual amino acids are different in this mutant compared to wild-type P450BM-3? What are some of
the known roles (in wild-type P450BM-3) of the changed amino acids?
b. What are the major products produced by the mutant when it is given noscapine as a substrate? (Hint:
Find the ‘supplementary material’ for the article for the best data.)
c. What is its ‘coupling efficiency’. What does this mean? How did the authors measure it?
d. What is the paper’s overall goal for the study, and does this mutant achieve their goal? Is it the best
mutant at achieving their goal?
e. Which of the human P450s tested in the Fang et al. paper is the VA mutant most like in terms of products
that it forms?

  1. In the Fang et al. article, they use a ‘dual-activity’ incubation system.
    a. What are the two activities, and why are both parts required to get the desired products?
    b. For UGT1A1, which glucuronide metabolites (C1, C3, or C4) was it capable of producing?
    c. In figure 6 panel A, it is shown that the products formed in the presence of NADPH but not GSH are
    different than those formed from the presence of both, and that the products formed in the presence of GSH
    but not NADPH are different that either of the other two sets of products. What does all of this tell us about
    the pathway generating the metabolites?
    d. Stare in awe at Figure 8 in all its glory. What is the difference between structure I and structure X? How
    do compounds VIII and IX relate to each other?

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