Team:Imperial College London/Drylab/Enzyme/Analysis
From 2009.igem.org
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- | Law of mass action assumptions: | + | <b>Law of mass action assumptions:</b> |
- | + | *Free diffusion | |
- | + | *Free unrestricted molecular motion | |
- | + | <br> | |
- | Michaelis-Menten assumption: | + | <b>Michaelis-Menten assumption:</b> |
- | + | *There is a quasi-steady state of [ES] , ie: , during which the enzyme is used for the reaction. This means that the rate of formation of ES complex is equal to the rate of dissociation of ES complex. | |
- | + | *<math> \begin{align} | |
- | + | K_M^{\prime} \ &\stackrel{\mathrm{def}}{=}\ \frac{k_3}{k_2 + k_3} K_M = \frac{k_3}{k_2 + k_3} \cdot \frac{k_{2} + k_{-1}}{k_{1}}\\ | |
+ | k_{cat} \ &\stackrel{\mathrm{def}}{=}\ \dfrac{k_3 k_2}{k_2 + k_3} | ||
+ | \end{align} </math> | ||
+ | *from the above assumptions, we know that | ||
{{Imperial/09/TemplateBottom}} | {{Imperial/09/TemplateBottom}} |
Revision as of 04:33, 22 September 2009
- Overview
- The model
- Simulations
Assumptions
Enzymatic assumptions:
- The enzyme is specific only for the substrate and not for any other chemicals.
- Only one enzyme, our enzyme of interest is present and participating in the reaction.
- There is negligible formation of product without the enzyme.
- The rate of enzymatic activity remains constant over time because:
- There is no co-operativity of the system. Binding of substrate to one enzyme binding site doesn't influence the affinity or activity of an adjacent site.
- No allosteric regulations from either the product or the substrate.
- There is no product inhibition of the enzyme.
- Total [E] does not change with regards to time. Enzyme will not be used up nor degraded over time, thus [E] at the beginning of the reaction will be the same as [E] at the end of reaction.
- The reaction catalysed is irreversible
- [S] >> [E], such that the free concentration of substrate is very close to the concentration I added. This also ensures a constant substrate concentration throughout the assay. This allows easy determination of [E].
- For which
<math> \begin{align} E + S \underset{k_{2}}{\overset{k_{1}} {\begin{smallmatrix}\displaystyle\longrightarrow \\ \displaystyle\longleftarrow \end{smallmatrix}}} ES \overset{k_3} {\longrightarrow} E + P \end{align}</math>
, k1>>k2
Law of mass action assumptions:
- Free diffusion
- Free unrestricted molecular motion
Michaelis-Menten assumption:
- There is a quasi-steady state of [ES] , ie: , during which the enzyme is used for the reaction. This means that the rate of formation of ES complex is equal to the rate of dissociation of ES complex.
- <math> \begin{align}
K_M^{\prime} \ &\stackrel{\mathrm{def}}{=}\ \frac{k_3}{k_2 + k_3} K_M = \frac{k_3}{k_2 + k_3} \cdot \frac{k_{2} + k_{-1}}{k_{1}}\\ k_{cat} \ &\stackrel{\mathrm{def}}{=}\ \dfrac{k_3 k_2}{k_2 + k_3} \end{align} </math>
- from the above assumptions, we know that