For future tests of the project in "Gut on a Chip" system, it is necessary to establish the optimal concentrations of nitrates needed to find the minimum concentration of the soluble transducer (sgp130) to inhibit transignaling.
For the prediction of the secreted concentrations of sgp130, a mathematical model was developed. This model relates the expressed protein concentration, secreted protein concentration, secretion rate, nitrate concentration and the rate of cell proliferation, where the cell line used needs IL6 to grow.
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Revision as of 21:20, 17 October 2018
Mathematical Model
Molecular approach
Simplified Model Of The Molecular Approach
Simbiology model
Using Matlab’s 2018 Simbiology tool we designed the model presented on the top; where C1 is the complex formed by (IL6/sIL6R) and C2 is the complex formed by (IL6/sIL6R/sgp130)
Differential Equations Model
A differential equations model is proposed where it is established that the proliferation of the HEPG2 cell line is a function of the concentration of nitrates, IL6, sIL6, sgp130, as well as, of the IL6 / sIL6 and IL6 / sIL6 / sgp130 complexes
Simbiology model
It is based on the hypothesis that by increasing the concentration of nitrates, the ammount of NsrR protein bound to DNA will decrease. This will lead to an increase in the levels of sgp130 synthesized that will bind to IL6/IL6R complex and thus, a decrease in the proliferation of the HEPG2 cell line will occurred. As can be seen in the figure presented in the left input parameters were obtained from literature.
References:
1. Crack JC, Svistunenko DA, Munnoch JT, Thomson AJ, Hutchings MI, Brun NEL. Differentiated, Promoter-specific Response of [4Fe-4S] NsrR DNA Binding to Reaction with Nitric Oxide*. In: The Journal of Biological Chemistry. ; 2016. doi:10.1074/jbc.M115.693192
2. Baran P, Hansen S, Waetzig GH, et al. The balance of interleukin (IL)-6, IL-6·soluble IL-6 receptor (sIL-6R), and IL-6·sIL-6R·sgp130 complexes allows simultaneous classic and trans-signaling. J Biol Chem. 2018;293(18):6762-6775. doi:10.1074/jbc.RA117.001163
3. Han S, Machhi S, Berge M, Xi G, Linke T, Schoner R. Novel signal peptides improve the secretion of recombinant Staphylococcus aureus Alpha toxinH35L in Escherichia coli. AMB Express. 2017;7. doi:10.1186/s13568-017-0394-1
4. Dumon-Seignovert L, Cariot G, Vuillard L. The toxicity of recombinant proteins in Escherichia coli: a comparison of overexpression in BL21(DE3), C41(DE3), and C43(DE3). Protein Expr Purif. 2004;37(1):203-206. doi:10.1016/j.pep.2004.04.025