5. In class we described two post-translational modifications which could be used to control enzymatic activity, phosphorylation and proteolysis. If we consider three enzymes which catalyze the same reaction, one of which is constitutively active (A), one which is activated by phosphorylation (B) and another which is activated by proteolysis (C), which enzyme, a. Requires the least energy to activate? b. Requires the least energy to inactivate?

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Chapter1: The Human Body: An Orientation
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b. Double mutant cycles rely on the concept of additivity. Let's imagine a hypothetical pair of side-chains in
the protein Tst1, Y5 and T78. The side-chain of Y5 makes a hydrogen bond to the T78 side-chain which
contributes 1.5 kcal/mol to the free energy change of folding of Tst1. The Y5 side-chain also makes
another 0.5 kcal/mol of favorable interactions with the rest of Tst1. Meanwhile, the T78 sidechain only
interacts with the Y5 side-chain. If we assume that neither the Y5A or the T78A mutations alter the
conformation of Tst1, what are the AAG values (the change in the free energy change upon folding) for:
i. The Y5A mutant
ii. The T78A mutant
iii.
E
The Y5A, T78A double mutant
4. The crude diagram below is an example of a folding funnel for a single polypeptide chain. It depicts a folded
state ("N") as well as a misfolded state ("MF"). Redraw this folding funnel, extending it to include two additional
states, i) a native dimer, ii) an aggregate. Under these conditions, let's assume that half of the folded protein is
dimerized and the majority of the remainder is aggregated.
W
MF
N
5. In class we described two post-translational modifications which could be used to control enzymatic activity,
phosphorylation and proteolysis. If we consider three enzymes which catalyze the same reaction, one of which is
constitutively active (A), one which is activated by phosphorylation (B) and another which is activated by
proteolysis (C), which enzyme,
a. Requires the least energy to activate?
b. Requires the least energy to inactivate?
Transcribed Image Text:b. Double mutant cycles rely on the concept of additivity. Let's imagine a hypothetical pair of side-chains in the protein Tst1, Y5 and T78. The side-chain of Y5 makes a hydrogen bond to the T78 side-chain which contributes 1.5 kcal/mol to the free energy change of folding of Tst1. The Y5 side-chain also makes another 0.5 kcal/mol of favorable interactions with the rest of Tst1. Meanwhile, the T78 sidechain only interacts with the Y5 side-chain. If we assume that neither the Y5A or the T78A mutations alter the conformation of Tst1, what are the AAG values (the change in the free energy change upon folding) for: i. The Y5A mutant ii. The T78A mutant iii. E The Y5A, T78A double mutant 4. The crude diagram below is an example of a folding funnel for a single polypeptide chain. It depicts a folded state ("N") as well as a misfolded state ("MF"). Redraw this folding funnel, extending it to include two additional states, i) a native dimer, ii) an aggregate. Under these conditions, let's assume that half of the folded protein is dimerized and the majority of the remainder is aggregated. W MF N 5. In class we described two post-translational modifications which could be used to control enzymatic activity, phosphorylation and proteolysis. If we consider three enzymes which catalyze the same reaction, one of which is constitutively active (A), one which is activated by phosphorylation (B) and another which is activated by proteolysis (C), which enzyme, a. Requires the least energy to activate? b. Requires the least energy to inactivate?
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