Coding

Part:BBa_K4810002

Designed by: Theodoros Desiniotis, Ioannis Stamelos, Aikaterini Spanou   Group: iGEM23_Athens   (2023-10-10)


Modified M13 pVIII protein for Perilic Aldehyde binding

M13 pVIII protein is the major coat protein of the M13 bacteriophage, found in 2700 copies in the surface of the phage accounting for 98% of its mass. pVIII is used for peptide display for the discovery of peptides able to bind with specific ligands (biopanning). Modifications in the pVIII protein can be used to create M13 bacteriophage-based colorimetric biosensors for the detection of specific molecules. The sensor's colorimetric output is a result of structural changes caused by ligand-pVIII protein interactions.[2]

Coding sequence of the protein is modified to express the specific hexapeptide (AEIPWT) in the N-terminal of the protein, exhibiting high binding affinity with the volatile organic compound "Perilic Aldehyde", found in decreased concentrations in the sebum of patients with Parkinson's disease.

The 23 amino acids of the N-terminal of pVIII protein undergo cleavage by a signal peptidase inside the bacterial cell, resulting in a functional protein of 50 amino acids. The specific modification occurred by deletion of the codons #2 to #5 of the functional protein and insertion of the respective codons of the desired peptide. Thus, the newly introduced peptide sequence is incorporated in the N-terminal of the protein.

The sequence of the specific peptide (AEIPWT) was determined by molecular docking simulations using Autodock Vina, opting for peptides with the highest affinity with Perilic Aldehyde and simultaneously the lowest affinity with other common organic compounds found in human sebum.

per.jpg


References

[1] - Lee, J., Warner, C., Jin, HE. et al. Production of tunable nanomaterials using hierarchically assembled bacteriophages. Nat Protoc 12, 1999–2013 (2017). https://doi.org/10.1038/nprot.2017.085

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None