Difference between revisions of "Part:BBa K3582304"

Latest revision as of 03:47, 28 October 2020

Inhibitory Sequence 1 for PfRH5-Basigin Grafted In Cyclotide

This composite biobrick is believed to synthesize the readily deliverable cyclotide drug that could inhibit the 5LGD interaction between the host protein (Basigin) and the parasite protein (PfRH5). Different components involved in this biobrick are described below:

1]BBa_K3582020: C intein is the modified version of the biobrick used by the Heidelberg 2014 team for circularization of the protein.
2]BBa_K2333015: Bsa 1 Reversed recognition site
3]BBa_K3582023(The cyclotide precursor): The gene coding for five loops of cyclotide Kalata B1 are inserted between the C Intein and the N intein so that it gets circularized after translation.
- Strep II tag(Inbuilt): This tag is inserted in loop 3 of the cyclotide construct to reduce the haemolytic activity of the cyclotide and ease of purification.
4]BBa_K3582303: Inhibitory peptide sequence 1 (IS1) grafted in loop 6 of the cyclotide structure to avoid the host-parasite protein interaction. This mutant sequence has a higher affinity for interaction as compared to wild type.
5]BBa_K3582021: Bsa 1 recognition site
6]BBa_K3582022: N intein is the modified version of the biobrick used by the Heidelberg 2014 team for circularization of the protein.

The inserted scars are basically the pieces of cyclotide precursor in whose loop the inhibitory peptide biobrick and strep II tag is grafted. The final translated protein product obtained using this biobrick is believed to act as a drug for preventing the CD36-PfEMP1 interaction.

Figure 1. Structure of Kalata B1 grafted with strep II tag and inhibitor based on homology modelling.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 112
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 143
Illegal BsaI.rc site found at 109

References

1] Gould, A., Ji, Y., Aboye, T. L., & Camarero, J. A. (2011, December). Cyclotides, a novel ultrastable polypeptide scaffold for drug discovery. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3330703/
2] DJ;, P. A. (n.d.). Cyclotides as grafting frameworks for protein engineering and drug design applications. Retrieved from https://pubmed.ncbi.nlm.nih.gov/23893608/
3] Camarero, J. A., & Campbell, M. J. (2019, April 19). The Potential of the Cyclotide Scaffold for Drug Development. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631875/

@@ Line 8: / Line 8: @@
 *3]BBa_K3582023(The cyclotide precursor): The gene coding for five loops of cyclotide Kalata B1 are inserted between the C Intein and the N intein so that it gets circularized after translation.
 **Strep II tag(Inbuilt): This tag is inserted in loop 3 of the cyclotide construct to reduce the haemolytic activity of the cyclotide and ease of purification.
-*4]BBa_k3582303: Inhibitory peptide sequence 1 (IS1) grafted in loop 6 of the cyclotide structure to avoid the host-parasite protein interaction. This mutant sequence has a higher affinity for interaction as compared to wild type.
+*4]BBa_K3582303: Inhibitory peptide sequence 1 (IS1) grafted in loop 6 of the cyclotide structure to avoid the host-parasite protein interaction. This mutant sequence has a higher affinity for interaction as compared to wild type.
 *5]BBa_K3582021: Bsa 1 recognition site
 *6]BBa_K3582022: N intein is the modified version of the biobrick used by the Heidelberg 2014 team for circularization of the protein.