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Topic: Michaelis-Menten Equation  (Read 11211 times)

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Offline ehFk

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Michaelis-Menten Equation
« on: May 15, 2009, 05:03:54 PM »
The laboratory I was supposed to perform on enzyme kinetics was cancelled and I was just given a set of data and I am not quite sure what the values mean with respect to the Michaelis-Menten equation.

Here are the resuts that I was given

[Substrate]   2 μM   4 μM     6 μM     8 μM     10 μM
 Slope #1   0.0711   0.1495   0.2585   0.3146   0.4151
 Slope #2   0.0874   0.1679   0.2222   0.2886   0.4111

As well as:
The kinetic assays were all read at 485 nm, enzyme volumes for each trial were constant (0.5 microL).

I am not quite sure what the slope value means with respect to the MM equation. I know that for the reciprocal of the MM equation the slope is equal to (Km/Vmax), and that the overall goal of the lab is to find Vmax and Kcat.

If anyone has an idea on how the slope I am given relates to the other variables, or any readings the can help me I would greatly appreciate it!

Offline sjb

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Offline Yggdrasil

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Re: Michaelis-Menten Equation
« Reply #2 on: May 16, 2009, 11:44:09 AM »
When performing enzyme kinetics measurements, you mix your enzyme and substrate then monitor the progress of the reaction over time (either the disappearance of substrate or appearance of product, let's say in this example that you are looking at the appearance of product).  If you plot the amount of product formed v. time, you will see a curve that goes up and slowly levels off as the reaction nears completion/equilibrium.  The Michaelis-Menten equation is concerned with the initial reaction rate.  So, when you look at the data, you want to look only at the first few data points of the [product] v. time graph, fit it to a line and record the slope.  Now you have one data point for your Michaelis-Menten curve (a concentration of substrate, [ S], and the corresponding initial reaction velocity, vo).  You would then repeat these kinetic measurements and determine vo for different concentrations of substrate while keeping other factors such as concentration of enzyme and temperature constant.

So, the slopes that you are given correspond to the initial reaction velocities (vo).
« Last Edit: May 16, 2009, 11:48:54 AM by Borek »

Offline ehFk

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Re: Michaelis-Menten Equation
« Reply #3 on: May 16, 2009, 04:00:24 PM »
Thank you very much Yggdrasil that helps out a lot!

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