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We are going to compare a lineweaver-Burk plot for regular kinetics and competit

ID: 482224 • Letter: W

Question

We are going to compare a lineweaver-Burk plot for regular kinetics and competitive inhibition. a. We showed in class [E]_Total = [E] + [ES] + [EI] and since we have equilibrium k_3[E][I] = k_3 [EI] so we can define the dissociation constant for the inhibitor as K_1 = k_3/k_3 so [EI] = [E][I]/K_1 Now substitute this back in equation (1) and solve for [E] b. As before, we set up steady state for [ES] d[ES]/dt = 0 = k 1 [E][S] - k - 1 [ES] - k2[ES] substitute your expression for [E] from part a. into this equation and solve for [ES] c. Rearrange to show that [ES] can be written as d. Now substitute this is your expression Rate = k2[ES] and compare your result to that of the traditional Lineweaver-Burk plot. In what ways is this plot different?

Explanation / Answer

Etotal = E+ES+EI

EI= [E] [I]/K

Etotal = E+ES+[E] [I]/K

Etotal = E(1+I/K)+ES

(Etotal –ES)(1+I/K) = E (1)

d(ES)/dt= K1[E][S]-K-1[ES]-K2[ES]=0

[ES] = K1[E] [S]/(K2+K-1)

From Eq.1 [ES] = K1{ Etotal- ES}(1+I/K)

[ES]= K1[Etotal]*(1+I/K) – ES*(1+I/K)

[ES] (2+I/K) = K1[Etotal] *(1+I/K)

ES = {K1[Etotal]*1+I/K)}/ (2+I/K)

R= K2[ES] = K2K1Etotal *(1+I/K)/(2+I/K)

1/R = (1+1+I/K)/(K2K1[Etotal]*(1+I/K)

1/R= 1/(K2K1[Etotal]*(1+I/K) + 1/K2K1[Etotal]

When a plot of 1/R vs 1/Etotal is drawn, the slope will be 1/K2K1 and intercept will an additional term 1/{K2K1total*(1+I/K)}

The difference is 1+I/K term in the intercept.

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