Part:BBa_K4143339

TEV Protease + T4GALA Encapsulin Targeting Peptide

Adding this sequence to the end of a protein serves to target it for sequestration inside the T4GALA encapsulin (BBa_K4143337). This is because each cargo protein in an encapsulin can be identified by the presence of a short C-terminal targeting peptide at its end. [1][2]

Usage and Biology

For our project, we aimed to use the targeting peptide to facilitate the encapsulation of an antimicrobial peptide (AMP) inside an encapsulin. The AMP sequence is described under part BBa_K4143336 and the encapsulin is characterized by part BBa_K4143337. To facilitate the encapsulation of our AMP, we attached the targeting peptide to the C-terminal end of the AMP.

AlphaFold Structural Characterization of AMP + Targeting Peptide

Because no structural information was available for our AMP + targeting peptide, we generated a tertiary structure using AlphaFold (Figure 1). Here, the targeting peptide is seen in the non-alpha-helical portion of the peptide, indicating it likely does not assume alpha helices or beta sheets for its secondary structure.

Figure 1: AlphaFold structural predictions for AMP HBCM2 + targeting peptide.

References

[1]T. W. Giessen and P. A. Silver, “Widespread distribution of encapsulin nanocompartments reveals functional diversity,” Nature Microbiology, vol. 2, no. 6, p. 17029, Mar. 2017, doi: 10.1038/nmicrobiol.2017.29.

[2]J. A. Jones, A. S. Cristie-David, M. P. Andreas, and T. W. Giessen, “Triggered reversible disassembly of an engineered protein nanocage,” bioRxiv, p. 2021.04.19.440480, Jan. 2021, doi: 10.1101/2021.04.19.440480.

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]