Team:Imperial College London/Modelling/Protein Display/Results and Conclusion

From 2010.igem.org

(Difference between revisions)
m
m
 
(13 intermediate revisions not shown)
Line 1: Line 1:
{{:Team:Imperial_College_London/Templates/Header}}
{{:Team:Imperial_College_London/Templates/Header}}
-
{| style="width:900px;background:#f5f5f5;text-align:justify;font-family: helvetica, arial, sans-serif;color:#555555;margin-top:25px;" cellspacing="20"
+
{{:Team:Imperial_College_London/Templates/ModellingHeaderD}}
 +
{{:Team:Imperial_College_London/Templates/ModellingDetectionHeader}}
 +
{|style="width:900px;background:#f5f5f5;text-align:justify;font-family: helvetica, arial, sans-serif;color:#555555;margin-top:5px;" cellspacing="20"
 +
|style="font-family: helvetica, arial, sans-serif;font-size:2em;color:#ea8828;"|Results and Conclusion
|-
|-
-
|Temporary sub-menu:<br/>
+
|The graph below represents the standard output of our model. It shows how the concentration of each of the species varies with time.  
-
<html>
+
 
-
<a href="https://2010.igem.org/wiki/index.php?title=Team:Imperial_College_London/Modelling/Protein_Display/Objectives"><b>Objectives</b></a>; <a href="https://2010.igem.org/Team:Imperial_College_London/Modelling/Protein_Display/Detailed_Description"><b>Detailed Description</b></a>; <a href="https://2010.igem.org/Team:Imperial_College_London/Modelling/Protein_Display/Parameters_and_Constants"><b>Parameters & Constants</b></a>; <a href="https://2010.igem.org/Team:Imperial_College_London/Modelling/Protein_Display/Results_and_Conclusion"><b>Results & Conclusion</b></a>;<a href="https://2010.igem.org/Team:Imperial_College_London/Modelling/Protein_Display/Download_MatLab_Files"><b>Download MatLab Files</b></a>;
+
<div ALIGN=CENTER>
-
</html>
+
  {| style="width:604px;background:#e7e7e7;text-align:center;font-family: helvetica, arial, sans-serif;color:#555555;margin- top:5px;padding: 2px;" cellspacing="5";
-
|}
+
-
{|style="width:900px;background:#f5f5f5;text-align:justify;font-family: helvetica, arial, sans-serif;color:#555555;margin-top:25px;" cellspacing="20"
+
-
|style="font-family: helvetica, arial, sans-serif;font-size:2em;color:#ea8828;"|Surface Protein Model
+
-
|-
+
-
|<div ALIGN=CENTER>
+
-
  {| style="background:#e7e7e7;text-align:center;font-family: helvetica, arial, sans-serif;color:#555555;margin- top:5px;padding: 2px;" cellspacing="5";
+
  |-
  |-
  |[[Image:IC_Protein_display.png|600px]]
  |[[Image:IC_Protein_display.png|600px]]
  |-
  |-
-
  |<html>Graphs showing the simulation using [TEV]<sub>0</sub> = 4&times;10<sup>-4</sup> mol/dm<sup>3</sup>. The graph on the right hand-side<br/> bottom shows that the AIP threshold (red line) is reached after 22 s.</html>
+
  |<html>Graphs showing the simulation of the enzymatic reaction for each of the species, with initial concentration of the enzyme - [TEV]<sub>0</sub> = 400 nM. The graph on the bottom right hand-side shows that the receptor activation threshold (red line) is reached by AIP after 11 s!</html>
  |}
  |}
</div>
</div>
Line 23: Line 20:
<ul>
<ul>
<li>Changing initial concentration of TEV</li>
<li>Changing initial concentration of TEV</li>
-
Whether the threshold concentration of AIP is reached is highly dependent on the initial concentration of TEV. The smallest initial concentration of TEV, [TEV>]<sub0</sub>, for which the threshold is reached is 6.0&times;10<sup>-6</sup>mol/dm<sup>3</sup>. On the grap below it can be seen that the optimal [TEV]<sub>0</sub> is a concentration higher than 10<sup>-4</sup>mol/dm<sup>3</sup>, which corresponds to the threshold being reached within 1.5 minutes.
+
Whether the threshold concentration of AIP is reached is highly dependent on the initial concentration of TEV. On the graph below it can be seen that the optimal [TEV]<sub>0</sub> is a concentration higher than 10nM, which corresponds to the threshold being reached within 9 minutes.
<br />
<br />
</html>
</html>
Line 29: Line 26:
  {| style="background:#e7e7e7;text-align:center;font-family: helvetica, arial, sans-serif;color:#555555;margin- top:5px;padding: 2px;" cellspacing="5";
  {| style="background:#e7e7e7;text-align:center;font-family: helvetica, arial, sans-serif;color:#555555;margin- top:5px;padding: 2px;" cellspacing="5";
  |-
  |-
-
  |[[Image:IC_AIP_Threshold_concentration.PNG|450px]]
+
  |[[Image:IC_AIP_threshold.png|600px]]
  |-
  |-
-
  |Graph showing when threshold AIP concentration is reached<br/> (for different initial TEV concentrations). Notice log-log scale.
+
  |Notice log-log scale.
  |}
  |}
</div>
</div>
Line 37: Line 34:
<li>Changing the production rate</li>
<li>Changing the production rate</li>
One order of magnitude change in the production rate results in at least 50s delay of the AIP concentration reaching the threshold concentration.<br/><br/>
One order of magnitude change in the production rate results in at least 50s delay of the AIP concentration reaching the threshold concentration.<br/><br/>
-
<li>Changing production rate</li>
 
-
Changing the production rate influences the time duration of the AIP concentration above the threshold level. The higher it is, the shorter the receptor will be activated (at extreme values, AIP concentration does not reach the threshold). However, the production rate has not much influence on how fast the threshold will be reached.<br/><br/>
 
-
<li>Changing control volume</li>
 
-
Our model is extremely sensitive to this factor. One order of magnitude change in CV results in several orders of magnitude change in AIP concentration. Hence, special care should be taken in determination of this value. If the model is to be compared with the experimental results, the CFU/ml has to be the same as the one used in the model. Otherwise, the CV has to be readjusted.
 
</ul>
</ul>
<br/>
<br/>
<b>Risk of False positives</b><br/>
<b>Risk of False positives</b><br/>
-
It was pointed out that we should assess the risk of false positive activation of the receptor. We are particularly concerned  about the display protein not binding to the cell wall, but instead diffusing into the extra-cellular environment.
+
It was pointed out that we should assess the risk of false positive activation of the receptor, but only areas of research were determined. We are particularly concerned  about the display protein not binding to the cell wall, but instead diffusing into the extra-cellular environment and maybe attaching to the receptor.
-
In order to be able to assess the risk of false positives, we need to do further research into the affinity of AIP with attached linker and transmembrane proteins for the receptor as compared to the affinity of the AIP itself for the receptor.
+
In order to be able to assess the risk of false positives, we need to do further research into the affinity of AIP with the attached linker and trans-membrane proteins for the receptor as compared to the affinity of the AIP itself for the receptor.
<br />
<br />
This paper <a href="http://jb.asm.org/cgi/content/full/186/10/3078">[1]</a> might have some information on affinity comparison.
This paper <a href="http://jb.asm.org/cgi/content/full/186/10/3078">[1]</a> might have some information on affinity comparison.
-
We need to know how proteins are being transported from intracellular to transmembrane space. Understanding this concept could give us an idea of what could go wrong.
+
We need to know how proteins are being transported from intracellular to trans-membrane space. Understanding of this concept could give us an idea of what could go wrong and on what modelling would need to focus on.
<br/>
<br/>
-
<h2>References</h2>
+
</html>
-
<ol>
+
|-
 +
|style="font-family: helvetica, arial, sans-serif;font-size:2em;color:#ea8828;" align="right"|[[Team:Imperial_College_London/Modelling/Protein_Display/Parameters_and_Constants | Click here for the constants of this model...]]
 +
|}
 +
{|style="width:900px;background:#f5f5f5;text-align:justify;font-family: helvetica, arial, sans-serif;color:#555555;margin-top:5px;" cellspacing="20"
 +
|style="font-family: helvetica, arial, sans-serif;font-size:2em;color:#ea8828;"|References
 +
|-
 +
|<html><ol>
<li>Knutsen, E., Ween, O. & Havarstein, L. (2003) Two Separate Quorum-Sensing Systems Upregulate Transcription of the Same ABC Transporter in Streptococcus pneumoniae. Journal of Bacteriology. [Online] 186(10), 3078-3085. Available from: http://jb.asm.org/cgi/reprint/186/10/3078 [Accessed 1st September 2010]</li>
<li>Knutsen, E., Ween, O. & Havarstein, L. (2003) Two Separate Quorum-Sensing Systems Upregulate Transcription of the Same ABC Transporter in Streptococcus pneumoniae. Journal of Bacteriology. [Online] 186(10), 3078-3085. Available from: http://jb.asm.org/cgi/reprint/186/10/3078 [Accessed 1st September 2010]</li>
</ol>
</ol>
</html>
</html>
|}
|}

Latest revision as of 03:21, 28 October 2010

Modelling Overview | Detection Model | Signaling Model | Fast Response Model | Interactions
A major part of the project consisted of modelling each module. This enabled us to decide which ideas we should implement. Look at the Fast Response page for a great example of how modelling has made a major impact on our design!
Objectives | Description | Results | Constants | MATLAB Code
Results and Conclusion
The graph below represents the standard output of our model. It shows how the concentration of each of the species varies with time.
IC Protein display.png
Graphs showing the simulation of the enzymatic reaction for each of the species, with initial concentration of the enzyme - [TEV]0 = 400 nM. The graph on the bottom right hand-side shows that the receptor activation threshold (red line) is reached by AIP after 11 s!


Sensitivity of our model

  • Changing initial concentration of TEV
  • Whether the threshold concentration of AIP is reached is highly dependent on the initial concentration of TEV. On the graph below it can be seen that the optimal [TEV]0 is a concentration higher than 10nM, which corresponds to the threshold being reached within 9 minutes.

    IC AIP threshold.png
    Notice log-log scale.


  • Changing the production rate
  • One order of magnitude change in the production rate results in at least 50s delay of the AIP concentration reaching the threshold concentration.


Risk of False positives
It was pointed out that we should assess the risk of false positive activation of the receptor, but only areas of research were determined. We are particularly concerned about the display protein not binding to the cell wall, but instead diffusing into the extra-cellular environment and maybe attaching to the receptor. In order to be able to assess the risk of false positives, we need to do further research into the affinity of AIP with the attached linker and trans-membrane proteins for the receptor as compared to the affinity of the AIP itself for the receptor.
This paper [1] might have some information on affinity comparison. We need to know how proteins are being transported from intracellular to trans-membrane space. Understanding of this concept could give us an idea of what could go wrong and on what modelling would need to focus on.

Click here for the constants of this model...
References
  1. Knutsen, E., Ween, O. & Havarstein, L. (2003) Two Separate Quorum-Sensing Systems Upregulate Transcription of the Same ABC Transporter in Streptococcus pneumoniae. Journal of Bacteriology. [Online] 186(10), 3078-3085. Available from: http://jb.asm.org/cgi/reprint/186/10/3078 [Accessed 1st September 2010]