Part:BBa_K404111
Mouse guanylate kinase (mGMK)
| Guanylate kinase (mGMK) | |
|---|---|
| |
| BioBrick Nr. | K404111 BBa K404111 |
| RFC standard | RFC 25 |
| Requirement | pSB1C3 |
| Source | synthetic |
| Submitted by | [http://2010.igem.org/Team:Freiburg_Bioware FreiGEM 2010] |
Guanylate kinases (GMKs) are involved in the salvage pathway of mono-phosphorylated guanosine (GMP) nucleosides to GDP therefore being essential in nucleotide maturation (Stolworthy & Black 2001). By introducing transgenic thymidine kinases (TKs) into tumor cells, a bottleneck occurs by overexpression of mono-phosphorylated intermediates. To overcome the accumulation of these non-toxic molecules, Willmon, Krabbenhoft, & Black (2006) fused the herpes simplex virus thymidine kinase (HSV-TK) to the guanylate kinase from M. musculus and demonstrated enhanced tumor killing in vitro.
Figure 1: Crystal structure (Sekulic et al. 2002) of mouse guanylate kinase in complex with GMP and ADP (PDB: 1LVG).
The iGEM team Freiburg_Bioware provides the mGMK BioBrick part within the ‘Virus Construction Kit’ for therapeutical applications. Producing viral particles with encapsidated single-stranded DNA containing the fusion gene mGMK_TK enhances the efficacy of the prodrug-activating system leading to cell death of targeted tumor cells
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 45
References
Sekulic, N. et al., 2002. Structural characterization of the closed conformation of mouse guanylate kinase. The Journal of biological chemistry, 277(33), 30236-43. Available at: http://www.ncbi.nlm.nih.gov/pubmed/12036965.Stolworthy, T.S. & Black, M.E., 2001. The mouse guanylate kinase double mutant E72Q/D103N is a functional adenylate kinase. Protein engineering, 14(11), 903-9. Available at: http://www.ncbi.nlm.nih.gov/pubmed/11742110.
Willmon, C.L., Krabbenhoft, E. & Black, M.E., 2006. A guanylate kinase/HSV-1 thymidine kinase fusion protein enhances prodrug-mediated cell killing. Gene therapy, 13(17), 1309-12. Available at: http://www.ncbi.nlm.nih.gov/pubmed/16810197.
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