The ITC traces for an experiment with various mutants of a key pathway kinase are shown below: Time (min) Time min) 30 40 Time (min) 10 20 50 10 50 0.10 no 00 000 ON 0.00 10 0.30 6.10 ucal 630 K70A 040 0.50 T75A 6. D193N ON 00 20 20 -20 40 40 calmolofinjectant kcal mol of injectant kcal mol of injectant -8.0 40 100 30 15 00 10 00 18 10 Molar Ratio OS Molar Ratio Molar Ratio Time (min) Time (min) Time (min) 020 4000 100 120 100 000 0.00 0.02 0.00 000 004 410 0.05 430 casc AN 015 -0.20 cal/sec 0.00 010 ucal/sec 40 F262W N255A F262A Bed 00- T ts N255A F262A F262W 18 M Time 75 Q305A H325A . molto Time (min) Time (min) 0.00 000 10 -610 020- 030 020 ucal ucalsec 9.30 -0.00 6.50- 040 Q305A H325A -0.00 00 00 20- 20- 40 kcal mor' of injectant kcal mol of injectant 40- 40 30 -100 20 00 05 10 15 20 25 30 35 Molar Ratio 00 05 10 Molar Ratio a. Which of the mutants appears to have the highest Ko? The lowest Ko? Be sure to explain why. b. How can you determine the stoichiometry of an interaction using ITC? What is the approximate stoichiometry of this interaction? c. How is the plot at the bottom of each panel derived from the collected data at the top of each panel? d. If a protein has a positive heat capacity of unfolding (Co), how do AHunfold and Sunfold change? Edit View Insert Format Tools Table 12nt Paragraph er Tv B I U The ITC traces for an experiment with various mutants of a key pathway kinase are shown below: Time (min) Time min) 30 40 Time (min) 10 20 50 10 50 0.10 no 00 000 ON 0.00 10 0.30 6.10 ucal 630 K70A 040 0.50 T75A 6. D193N ON 00 20 20 -20 40 40 calmolofinjectant kcal mol of injectant kcal mol of injectant -8.0 40 100 30 15 00 10 00 18 10 Molar Ratio OS Molar Ratio Molar Ratio Time (min) Time (min) Time (min) 020 4000 100 120 100 000 0.00 0.02 0.00 000 004 410 0.05 430 casc AN 015 -0.20 cal/sec 0.00 010 ucal/sec 40 F262W N255A F262A Bed 00- T ts N255A F262A F262W 18 M Time 75 Q305A H325A . molto Time (min) Time (min) 0.00 000 10 -610 020- 030 020 ucal ucalsec 9.30 -0.00 6.50- 040 Q305A H325A -0.00 00 00 20- 20- 40 kcal mor' of injectant kcal mol of injectant 40- 40 30 -100 20 00 05 10 15 20 25 30 35 Molar Ratio 00 05 10 Molar Ratio a. Which of the mutants appears to have the highest Ko? The lowest Ko? Be sure to explain why. b. How can you determine the stoichiometry of an interaction using ITC? What is the approximate stoichiometry of this interaction? c. How is the plot at the bottom of each panel derived from the collected data at the top of each panel? d. If a protein has a positive heat capacity of unfolding (Co), how do AHunfold and Sunfold change? Edit View Insert Format Tools Table 12nt Paragraph er Tv B I U