Difference between revisions of "Team:BIT-China/H2O2DecompositionModel"

 
(7 intermediate revisions by 2 users not shown)
Line 46: Line 46:
 
         #HQ_page td{
 
         #HQ_page td{
 
             vertical-align: inherit !important;
 
             vertical-align: inherit !important;
 +
        }
 +
 +
        figcaption {
 +
            color: #131313;
 
         }
 
         }
 
     </style>
 
     </style>
Line 91: Line 95:
 
         }
 
         }
 
     </style>
 
     </style>
    <script src="https://2018.igem.org/Template:BIT-China/js/base-loading?action=raw&ctype=text/javascript"></script>
+
 
 
     <script type="text/x-mathjax-config">
 
     <script type="text/x-mathjax-config">
 
         MathJax.Hub.Config({
 
         MathJax.Hub.Config({
Line 105: Line 109:
 
</head>
 
</head>
  
<body id="ibody" class="scoll_dis">
+
<body>
  
    <ul id="left-nav">
+
            <ul id="left-nav">
 
         <li>
 
         <li>
 
             <a>PROJECT</a>
 
             <a>PROJECT</a>
Line 123: Line 127:
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsFeedback">Feedback</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsFeedback">Feedback</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsOutput">Output</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsOutput">Output</a></li>
                 <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsInput">Input</a></li>
+
                 <li><a href="https://2018.igem.org/Team:BIT-China/Results">Results</a></li>
 
             </ul>
 
             </ul>
 
         </li>
 
         </li>
Line 131: Line 135:
 
             <ul>
 
             <ul>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/Model">Overview</a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/Model">Overview</a></li>
                 <li><a href="https://2018.igem.org/Team:BIT-China/FluorescentProbesModel">Fluorescent Probes Model </a></li>
+
                 <li><a href="https://2018.igem.org/Team:BIT-China/FluorescentProbesModel">Fluorescent Probe Model </a></li>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/H2O2DecompositionModel">H<sub>2</sub>O<sub>2</sub>
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/H2O2DecompositionModel">H<sub>2</sub>O<sub>2</sub>
 
                         Decomposition Model</a></li>
 
                         Decomposition Model</a></li>
  
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/roGFP2-Orp1MichaelisEquationModel">roGFP2-Orp1
 
                 <li><a href="https://2018.igem.org/Team:BIT-China/roGFP2-Orp1MichaelisEquationModel">roGFP2-Orp1
                         Michaelis equations Model</a></li>
+
                         Michaelis equation Model</a></li>
 
             </ul>
 
             </ul>
 
         </li>
 
         </li>
Line 192: Line 196:
 
         <div class="MD-title">
 
         <div class="MD-title">
 
             <a style="border-bottom-style: solid;text-decoration: none;color:#131313;">H<sub>2</sub>O<sub>2</sub>
 
             <a style="border-bottom-style: solid;text-decoration: none;color:#131313;">H<sub>2</sub>O<sub>2</sub>
                 Decomposition Model</a>
+
                 DECOMPOSITION MODEL</a>
 
         </div>
 
         </div>
  
Line 209: Line 213:
 
                     up a model to simulate
 
                     up a model to simulate
 
                     the intracellular H<sub>2</sub>O<sub>2</sub>. At the same time, this model will provide a base for
 
                     the intracellular H<sub>2</sub>O<sub>2</sub>. At the same time, this model will provide a base for
                     roGFP2-Orp1 Micheali
+
                     roGFP2-Orp1 Michealis
 
                     equation model.
 
                     equation model.
 
                 </p>
 
                 </p>
Line 226: Line 230:
 
                     some natural reduction molecular (like Glutathione) directly react with H<sub>2</sub>O<sub>2</sub>.
 
                     some natural reduction molecular (like Glutathione) directly react with H<sub>2</sub>O<sub>2</sub>.
 
                     And slow degradation
 
                     And slow degradation
                     is led by enzymatic degradation.(一篇参考文献)
+
                     is led by enzymatic degradation.<sup onmouseover="tooltip.pop(this, '#tip-1', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
 +
                    style="color:#38679a;">[1]</sup>
 
                 </p>
 
                 </p>
  
 
                 <figure class="MD-Fig MD-margin-toContentP">
 
                 <figure class="MD-Fig MD-margin-toContentP">
                     <img src="https://static.igem.org/mediawiki/2018/d/d3/T--BIT-China--iGEM2018-Modeling7.png">
+
                     <img src="https://static.igem.org/mediawiki/2018/9/92/T--BIT-China--modelingh202decompositiondig1chaizong.png">
                     <figcaption>Fig.1 The kinetic model for H2O2 degradation</figcaption>
+
                     <figcaption>Fig.1 The kinetic model for H<sub>2</sub>O<sub>2</sub> degradation</figcaption>
 
                 </figure>
 
                 </figure>
 
             </div>
 
             </div>
Line 255: Line 260:
  
 
                 <p class="MD-content-p">
 
                 <p class="MD-content-p">
                     The ODE model accounted for intra- and extracellular H<sub>2</sub>O<sub>2</sub> levels, and the
+
                     The ODE model accounted for intracellular and extracellular H<sub>2</sub>O<sub>2</sub> levels, and the
 
                     consumption of a finite
 
                     consumption of a finite
 
                     antioxidant capacity using three rates:
 
                     antioxidant capacity using three rates:
Line 262: Line 267:
 
                 <ul class="MD-content-p">
 
                 <ul class="MD-content-p">
 
                     <li>
 
                     <li>
                         1) The membrane permeability (k1).(三篇参考文献)
+
                         1) The membrane permeability (k1).<sup onmouseover="tooltip.pop(this, '#tip-2', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
 +
                        style="color:#38679a;">[2]</sup><sup onmouseover="tooltip.pop(this, '#tip-3', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
 +
                        style="color:#38679a;">[3]</sup><sup onmouseover="tooltip.pop(this, '#tip-4', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
 +
                        style="color:#38679a;">[4]</sup>
 
                     </li>
 
                     </li>
 
                     <li>
 
                     <li>
                         2) Intracellular slow degradation rate (k2).(一篇参考文献)
+
                         2) Intracellular slow degradation rate (k2).<sup onmouseover="tooltip.pop(this, '#tip-5', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
 +
                        style="color:#38679a;">[5]</sup>
 
                     </li>
 
                     </li>
 
                     <li>
 
                     <li>
 
                         3) Intracellular fast degradation rate (k3) dependent upon a finite capacity (g), e.g. NADPH
 
                         3) Intracellular fast degradation rate (k3) dependent upon a finite capacity (g), e.g. NADPH
                         levels in naive cells.(一篇参考文献)
+
                         levels in naive cells.<sup onmouseover="tooltip.pop(this, '#tip-6', {position:1, offsetX:-20, effect:'slide',hideDelay: 10})"
 +
                        style="color:#38679a;">[6]</sup>
 
                     </li>
 
                     </li>
 
                 </ul>
 
                 </ul>
Line 383: Line 393:
 
                     To verify the feasibility of the model and to confirm this dynamic process, we did some experiments
 
                     To verify the feasibility of the model and to confirm this dynamic process, we did some experiments
 
                     using our system's output, roGFP2-Orp1. The results show that the fluorescence ratio value
 
                     using our system's output, roGFP2-Orp1. The results show that the fluorescence ratio value
                     I405/I488, an oxidant condition indicator, decreased by time when S. cerevisiae exposed to
+
                     I405/I488, an oxidant condition indicator, decreased by time when <i>S. cerevisiae</i> exposed to
 
                     different concentrations of H<sub>2</sub>O<sub>2</sub>. This ratio's decreasing stands for the
 
                     different concentrations of H<sub>2</sub>O<sub>2</sub>. This ratio's decreasing stands for the
 
                     increasing of
 
                     increasing of
                     intercellular H2O2 concentration. It also stopped at a stable concentration at last, which's
+
                     intercellular H<sub>2</sub>O<sub>2</sub> concentration. It also stopped at a stable concentration at last, which's
 
                     correspond to our model's result.
 
                     correspond to our model's result.
 
                 </p>
 
                 </p>
Line 412: Line 422:
 
         </div>
 
         </div>
 
     </div>
 
     </div>
 +
    <div style="display:none;font-family: 'helveticaregular';">
 +
        <div id="tip-1">
 +
            <div class="column" style="width:230px;height: 100px;">
 +
                <a>Altıntaş A, Davidsen K, Garde C, et al. High-resolution kinetics and modeling of hydrogen peroxide degradation in live cells.[J]. Free Radical Biology & Medicine, 2016, 101:143-153.
 +
                </a>
 +
            </div>
 +
        </div>
  
 +
        <div id="tip-2">
 +
            <div class="column" style="width:230px;height: 100px;">
 +
                <a>Branco M R, Marinho H S, Cyrne L, et al. Decrease of H<sub>2</sub>O<sub>2</sub> plasma membrane permeability during adaptation to H<sub>2</sub>O<sub>2</sub> in Saccharomyces cerevisiae.[J] . Journal of Biological Chemistry, 2004, 279(8):6501
 +
                </a>
 +
            </div>
 +
        </div>
 +
 +
        <div id="tip-3">
 +
            <div class="column" style="width:230px;height: 100px;">
 +
                <a>Henzler T, Steudle E. Transport and metabolic degradation of hydrogen peroxide in Chara corallina: model calculations and measurements with the pressure probe suggest transport of H(2)O(2) across water channels.[J]. Journal of Experimental Botany, 2000, 51(353):2053-2066.
 +
                </a>
 +
            </div>
 +
        </div>
 +
        <div id="tip-4">
 +
            <div class="column" style="width:230px;height: 100px;">
 +
                <a>Bienert G P, Schjoerring J K, Jahn T P. Membrane transport of hydrogen peroxide[J]. Biochimica Et Biophysica Acta, 2006, 1758(8):994-1003.
 +
                </a>
 +
            </div>
 +
        </div>
 +
        <div id="tip-5">
 +
            <div class="column" style="width:230px;height: 100px;">
 +
                <a>Branco M R, Marinho H S, Cyrne L, et al. Decrease of H<sub>2</sub>O<sub>2</sub> plasma membrane permeability during adaptation to H<sub>2</sub>O<sub>2</sub> in Saccharomyces cerevisiae.[J]. Journal of Biological Chemistry, 2004, 279(8):6501.
 +
                </a>
 +
            </div>
 +
        </div>
 +
        <div id="tip-6">
 +
            <div class="column" style="width:230px;height: 100px;">
 +
                <a>Altıntaş A, Davidsen K, Garde C, et al. High-resolution kinetics and modeling of hydrogen peroxide degradation in live cells.[J]. Free Radical Biology & Medicine, 2016, 101:143-153.
 +
                </a>
 +
            </div>
 +
        </div>
 +
 +
 +
    </div>
 
     <!-- footer start -->
 
     <!-- footer start -->
 
     <div class="footer-all" style="margin-top:65px;">
 
     <div class="footer-all" style="margin-top:65px;">

Latest revision as of 00:44, 18 October 2018

In our experiment, we found it hard to measure the intracellular H2O2 concentration, so we controlled the external H2O2 concentration in culture to confirm our system' function. Although the H2O2 can entry the cell quickly through simple diffusion, we needed to build up a model to simulate the intracellular H2O2. At the same time, this model will provide a base for roGFP2-Orp1 Michealis equation model.

Firstly, in order to describe the process in cells more accurately, three reactions in this process are considered: diffusion, fast degradation and slow degradation. The fast degradation is caused by some natural reduction molecular (like Glutathione) directly react with H2O2. And slow degradation is led by enzymatic degradation.[1]

Fig.1 The kinetic model for H2O2 degradation

$$\frac{\mathrm{d_{\mathit{p}_{e}}} }{\mathrm{d} t}=k_{1}(p_{i}-p_{e})v$$

$$\frac{\mathrm{d_{\mathit{p}_{i}}} }{\mathrm{d}t}=k_{1}(p_{e}-p_{i})-(k_{2}+k_{3}\frac{g}{k_{g}+g})p_{i}$$

$$\frac{\mathrm{d_{\mathit{g}}} }{\mathrm{d} t}=-k_{3}(\frac{g}{k_{g}+g})p_{i}$$

The ODE model accounted for intracellular and extracellular H2O2 levels, and the consumption of a finite antioxidant capacity using three rates:

  • 1) The membrane permeability (k1).[2][3][4]
  • 2) Intracellular slow degradation rate (k2).[5]
  • 3) Intracellular fast degradation rate (k3) dependent upon a finite capacity (g), e.g. NADPH levels in naive cells.[6]

We reviewed the above literature and combined the results of the wet experiment, as shown in Fig.3, to determine the parameters required for the model simulation, listed in the table below.

Table 1. Description of Variables Used in The Kinetic Model.

Variable Explanation Value
k1 Membrane Permeability 11
k2 Slow Degradation Rate 8
k3 Fast Degradation Rate 55
G Finite Capacity Substance 60
pi Intracellular/Cytosolic [H2O2] /
pe Extracellular [H2O2] /
V Ratio of Cell Volume to Media 0.005

We used MATLAB to simulate the kinetics of extracellular and intracellular H2O2 changes over time. (Fig. 2)

Fig.2 Simulation result

To verify the feasibility of the model and to confirm this dynamic process, we did some experiments using our system's output, roGFP2-Orp1. The results show that the fluorescence ratio value I405/I488, an oxidant condition indicator, decreased by time when S. cerevisiae exposed to different concentrations of H2O2. This ratio's decreasing stands for the increasing of intercellular H2O2 concentration. It also stopped at a stable concentration at last, which's correspond to our model's result.

Fig.3 Fluorescence ratio value changes with time

In this model, we successfully simulated the kinetics of extracellular and intracellular H2O2 changes over time. Furthermore, the dynamic process of fluorescence ratio value by time was detected by using roGFP2-Orp1 green fluorescent protein in wet experiment, and the feasibility of the model was further verified.