Part:BBa_K4850006

Rigid Protein Linker A(EAAAK)₂A

A(EAAAK)₂A is a specific type of rigid protein linker used for genetically fusing functional protein domains. It can maintain a fixed distance between functional domains while preserving their individual functions. This is achieved through the adoption of an alpha-helical structure, which is renowned for its rigidity and stability, attributed to the closely packed backbone structure and the presence of intra-segment hydrogen bonds. The A(EAAAK)₂A protein linker was originally designed by Arai et al. (2001), and it features an empirical sequence denoted as A(EAAAK)_nA, where n typically ranges from 2 to 5.

Usage and Biology

The NUS-Singapore 2023 iGEM team frequently employed this linker in their gene circuits when there was a need to connect two protein coding sequences to create a fusion protein. The part that made use of this linker include:

• BBa_K4850008: GFP-sacB Negative Selection Marker (Composite Part)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 7
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Arai, R., Ueda, H., Kitayama, A., Kamiya, N., & Nagamune, T. (2001). Design of the linkers which effectively separate domains of a bifunctional fusion protein. Protein Engineering Design & Selection, 14(8), 529–532. https://doi.org/10.1093/protein/14.8.529

[2] Chen, X., Zaro, J. L., & Shen, W. (2013). Fusion protein linkers: Property, design and functionality. Advanced Drug Delivery Reviews, 65(10), 1357–1369. https://doi.org/10.1016/j.addr.2012.09.039