Part:BBa_K4905007
Elastin-Like Polypeptide Triblock with Leucine Zippers
Information
This part is made up of the basic parts: Leucine zipper Z1 (BBa_K4905004), Leucine zipper Z2 (BBa_K4905005), Elastin-like polypeptide sequence A[60]I[60] (BBa_K4905001), and Elastin-like polypeptide sequence A[40]I[60] (BBa_K4905002). It was used by the TU Eindhoven 2023 team to form a hydrogel outside as well as inside e.coli BL21 cells.
The construct consists of Elastin-like Polypeptides (ELPs) and two different leucine zippers that have affinity for each other. In general, ELPs have hydrophilic and hydrophobic domains that exhibit reversible phase separation behavior that is temperature dependent. As shown in figure ..., this construct has a hydrophilic region in the middle and a hydrophobic region on each side of it. On both ends, the leucine zippers Z1 and Z2 are located for stronger interactions between the ELPs. This allows them to be used in the formation of a reversible hydrogel at a temperature of 37 °C.
Figure 1: Schematic representation of the composite part, an Elastin-Like Polypeptide with leucine zippers on the ends
Application
In the TU Eindhoven 2023 team project, the ELPs are, in the end, expressed inside e.coli BL21 cells. The hydrogel will be formed with electrostatic and hydrophobic interactions. At a temperature below 37 °C, the hydrophobic parts are surrounded by water molecules. This causes a low entropy state. When the temperature increases over 37 °C, these water molecules go into the bulk water phase what gains the solvent entropy. This makes it possible to form interactions with other ELP molecules [1].
When the hydrogel is formed inside e.coli BL21 cells, it prevents the cell from dividing. However, the cells remain functional. So they can still be used to express therapeutic agents, like Interleukin 10 in the project of the TU Eindhoven 2023 team.
Sequence and Features
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1982
Illegal XbaI site found at 99 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1982
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1982
Illegal XhoI site found at 1999 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1982
Illegal XbaI site found at 99 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1982
Illegal XbaI site found at 99
Illegal NgoMIV site found at 156
Illegal NgoMIV site found at 336
Illegal NgoMIV site found at 426
Illegal NgoMIV site found at 606
Illegal NgoMIV site found at 2928
Illegal NgoMIV site found at 3105
Illegal NgoMIV site found at 3195 - 1000COMPATIBLE WITH RFC[1000]
Results
References
[1] Despanie, J., Dhandhukia, J. P., Hamm-Alvarez, S. F., & MacKay, J. A. (2016). Elastin-like polypeptides: Therapeutic applications for an emerging class of nanomedicines. Journal of Controlled Release, 240, 93–108. https://doi.org/10.1016/j.jconrel.2015.11.010
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