Question

Biochemistry question please help

Congratulations! You have just started working in a brand spanking new biochemistry lab with some very fancy equipment to accurately measure enzyme activities. There is only one problem...You work for an absent minded Investigator who cannot keep track of his inhibitors. He has given you a series of inhibitors to certain enzymatic activity and asked you to sort through them. He would like to know a) What type of inhibitor is each compound (Competitive, non-competitive, uncompetitive or mixed) and b) what is the Ki for each inhibitor You have run the experiments and the data is in the table below. Please transfer this data into an Excel file and plot the data both as v vs. [S] curves and double-reciprocal plots. Determine the type of inhibitor and calculate the Ki for each. Please upload your Excel file with the plots and your answers incorporated S] (mM) 0.2 0.25 0.333 0.5 X 30uM 5.56 6.67 8.33 Y@4mM Control 16.67 20 24.98 33.33 50 66.67 71.4 76.92 80 83.33 6.25 7.69 10 14.29 25 40 45.45 52.63 57.14 62.5 16.67 22.22 23.81 25.64 26.67 27.77 12.5 14.29 16.67 18.18 18.52 18.87 19 19.23 Z 0.2mMM 8.89 10.81 13.78 19.05 30.77 44.44 48.78 54.06 57.14 60.6 2.5 3.33 4

Answer 1

Lineweaver-Burk Inversion 0.20 ● 1 /Vo control Use the equation for a straight line: y-ax+Ib 9.99E-03*x+9.99E-03 slope y-intercept 0.03*x+9.98E-03 0.15 0.0299x+0.03 9.98E-03% + 0.05 1/No Z Z 0.2mM 0.02% + 0.0125 0.10 Plot 1/v (y) versus 1/[S] (x) slope- 0.05 x-intercept =-1/KM y-intercept = 1/Vmax 1/[S] 0.00 2.000 0.000 2.000 4.000

Use the equation on the plot Control Y-0.0098x + 0.0099 y 0.03x0.0099 y = 0.0299x + 0.03 1/Vmax- Vmax- 1/Vmax- 0.0099 0.0099 0.03 Vmax- 101.010 mM/min 101.0101 mM/min Vmax - 33.3333uM/sec km/Vmax Km (-- 0.009 0.03 ki/Vmax- 0.0299 0.909 mM Ki(-)s 3.030 mM 1.00 Competitive inhibitor increase Ki do not affect Vmax Non Competitive inhibitor do not change Km, but decreases Vmax