Coding
sgp130

Part:BBa_K2760002:Design

Designed by: iGEM18_TecMonterrey_GDL   Group: iGEM18_TecMonterrey_GDL   (2018-10-08)

gp130: (soluble human gp130, transducer for IL-6 complex)

This gene codes for the soluble human IL-6 transducer signaling glycoprotein 130, which is the beta chain of the IL-6 receptor complex. When gp80 and gp130 join that a high affinity complex for IL-6 is formed, thus binding with IL-6 occurs [1]. Studies have shown that this soluble form of gp130 is an inhibitor to trans-signaling events in the human body, via binding with the dimer of IL-6 and soluble gp80 (receptor, which circulates in the extracellular media [1]. This piece is meant to be used in a system to antagonically reduce trans-signaling. The inhibition of trans-signaling with novel drugs have been previously proposed to treat depressive patients [2], thus this piece was used for the design of Lactobachill (iGEM18_TecMonterrey_GDL), a proposed psychobiotic to be used as an adjoint in the treatment and prevention of depression.

Interleukin 6 (IL-6) is a multifunctional but majorly a proinflammatory cytokine within the IL-6 family. IL-6 signalization occurs via glycoprotein 130 (gp130, beta chain, of 130kDa) and the membrane-bound or soluble IL-6 receptor (gp80, alpha chain, of 80kDa), these piece is meant to unite with gp80, to form the high affinity complex in order to bind with IL-6 [3].

Using a molecular weight calculator online the protein weight for the amino acid sequence resulted to be 34.86 kilodaltons [4].

    • The sequence is codon optimized for E. coli and L. rhamnosus

The sequence was obtained from Seegert et al., the optimized nucleotide encoding sgp130 (2012) [5] then other articles were taken as reference to verify it was the desired sequence, from Xu et. al [6], Wolf et. al [7] and Wolf, Waetzig, et. al [8].

References:

[1] Jostock, T., Müller, j., Ozbek, S., Atreya, S., Blinn, G., Voltz, N., Fischer, M., Neurath, M., Rose-John, S. (2001). Soluble gp130 is the natural inhibitor of soluble interleukin-6 receptor transsignaling responses. Eur. J. Biochem 268, 160-167. FEBS.

[2] Fonseka, T., Melntyre, R., Soczynska, J., Kennedy, S. (2015). Novel investigation drugs targeting IL-signaling for the treatment of depression. Informa UK. 10.1517/13543784.2014.99833 ISSN 1354-3784, e-ISSN 1744-7658

[3] Müller-Newen, G., Küster, A., Hemmann, U., Keul,R., Horsten, U., Martens, A., Graeve, L., Wijdenes, J., Heinrich, P. (1998) Soluble IL-6 Receptor Potentiates the Antagonistic Activity of Soluble gp130 on IL-6 Responses. The Journal American of Association Immunology. ISSN: 1550-6606.

[4]. Molecular weight calculator. Science Gateway. https://www.sciencegateway.org/tools/proteinmw.htm

[5] Seegert, D., Wätzig, K., Rahaus, N., Daecke, J., Rose-John, S. (2012). Optimized nucleotide sequences encoding sgp130. US 8,206,984 B2.

[6] Xu, Y.,.Kershaw, N., Luo, C.,Soo, P., Pocock, M., Czabotar, P., Hilton, D., Nicola, N., .Garrett, T., Zhang, J. (2010) Crystal Structure of the Entire Ectodomain of gp130. The Journal of Biological Chemistry Vol 285. NO 28,pp21214-21218. [7] Wolf, J., Waetzig, G., Chalaris, A., Reinheimer, T., Wege, H., Rose-John, S., and Garbers, C. (2016). Different Soluble Forms of the Interleukin-6 Family Signal Transducer gp130 Fine-Tune the Blockade of Interleukin-6 Trans-Signaling. JBC: http://www.jbc.org/cgi/doi/10.1074/jbc.M116.718551

[8] J. Wolf, G.H. Waetzig, T.M. Reinheimer, J. Scheller, S. Rose-John, C. Garbers, A soluble form of the interleukin-6 family signal transducer gp130 is dimerized via a C-terminal disulfide-bridge resulting from alternative mRNA splicing, Biochemical and Biophysical Research Communications (2016), doi: 10.1016/j.bbrc.2016.01.127