a and b please EXAM #3-100 pts, 4 Pages 6. A certain enzyme with as quaternary s

Question

a and b please

EXAM #3-100 pts, 4 Pages 6. A certain enzyme with as quaternary structure contains one active site per 85 kDa subunit. A series of enzyme assays with (and without) an inhibitor are carried out. Each enzyme assay contained 3.3 ug of enzyme. The enzyme assay data is plotted below. Answer the questions that follow. Clearly show all of your work. 30 ys 4.1x (2.9 R2 = 0.996 SS 29 E25 + Inhibitor (20 ?M) a 20 E C 15 1.9x + 2.9 R2:0.9902 No additions m,? 0 6 8 1/IS] (mM) DEFINE Kea then use the data above to CALCULATE the value of Kcat (min2) in the absence of inhibitor. en then use the data above to CALCULATE the value of ka (min") in the absence of inhibitor ? de vaI (b) DEFINE Km AND then calculate its value (in mM) using the data above. Slo vm

Explanation / Answer

(a)

From the plot

intercept at y-axis = 1/Vmax = 2.5

Vmax = 1/2.5 = 0.4 umol/L.min

Kcat = Vmax/[Et]

[Et] = 3.3 ug/85000

= 4 x 10^-5 umol

So, Kcat = 0.4/4 x 10^-5

= 10,000 min-1

(b)

Km = -1/intercept on x-axis

            = -(-1/1 mM)

            = 1 mM

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