4. What does enzyme kinetics study? What is Vo, km, Vmax, Kcat, respectively? If you plot...
A.Your summer research project is to study the kinetics of a newly discovered enzyme. You have already identified the substrate and optimal reaction conditions. Propose how you would set up experiments to characterize the maximum velocity (Vmax) and Michaelis constant (KM) of this enzyme, assuming that it will display Michaelis-Menten kinetics. B. How would you compare the specificity of the enzyme for its natural substrate versus its specificity for a chromogenic substrate analog that was synthesized in the lab
Does increasing enzyme concentration changes the value of Vmax, Km and Kcat? Why?
16. At right is a graph obtained from a series of enzyme kinetics assays. The Vmax for this enzyme and substrate is 4.5 uM/s. 5 4.5 4 a) What is the KM? KM: v. (mM/s) 3.5 3 2.5 2 1.5 1 0.5 b) If a pure non-competitive inhibitor was added to the assays, what would the resulting kinetics curve be like? Give a Km and Vmax in the presence of the inhibitor (write them below) and draw an appropriate curve...
1. Show, using the Michaelis-Menten equation, that when [S] >>> Km, vo = Vmax. Show, using the M-M equation that when [S] <<<Km, vo =[S][Et]kcat/Km. 2. What is Vmax? Provide both a mathematical and written description of Vmax? How can Vmax be experimentally altered? How can we use Vmax to determine the turnover number (kcat) of an enzyme-catalyzed reaction? What is the major challenge of determining Vmax from an Michaelis-Menten plot?
Values for an Enzyme and a substrate are: Km=4 uM and kcat=20/min. For an experiment where [S]=6mM, it was found that Vo=480nM/min. What was the enzyme concentration? Give your answer in nM. Using the same kcat and Km as the previous question, if [Et]=0.5uM gives a Vo=5 uM/min, what wat the [S]? Give your answer in uM. reaction is run with the kcat=20/min and the Km=4uM. Use the enzyme concentration from question 1 above. A very strong inhibitor is added creating...
16. At right is a graph obtained from a series of enzyme kinetics assays. The Vmax for this enzyme and substrate is 4.5 uM/s. a) What is the KM? 5 4.5 4 3.5 3 2.5 2 KM: 3mm V. (mM/s) 1.5 1 b) If a pure non-competitive inhibitor was added to the assays, what would the resulting kinetics curve be like? Give a Km and Vmax in the presence of the inhibitor (write them below) and draw an appropriate curve...
D-Lactose is the substrate for B-galactosidase. Given Vo = kcat [Et] [S]/km + [S], calculate [S], when Km = 4.0 nM, V. = 10.5 M s', kcat = 500 s, and [Et] = 40 uM Calculate the catalytic efficiency. Below is a double-reciprocal plot for an enzyme reaction in the absence and presence of of inhibitor. Give the equation for the line. Calculate Vmax and Km for the enzyme and enzyme plus inhibitor. Which type of inhibition is apparent. 0.10...
Consider the enzyme-catalyzed reaction with Vmax=164 (μmol/L)min−1 and KM=32μmol/L. Part A If the total enzyme concentration was 6 nmol/L, how many molecules of substrate can a molecule of enzyme process in each minute? Express your answer to three significant figures. kcat kcat = 2.73×104 min−1 Part B Calculate kcat/KM for the enzyme reaction.
An enzyme kinetics experiment is performed with the enzyme Imaginase, which converts Horses into Unicorns with a known Kcat of 300 s-. In an experiment with (Imaginase) at 40 nM and (Horses) at 10 uM, the researchers find that Vo = 4 uM/s. What is the Km of Imaginase for its substrate Horses?
How do competitive inhibitors affect the KM and Vmax of an enzyme? Draw a plot of velocity as a function of substrate concentration, both with and without inhibitor added.