Difference between revisions of "Part:BBa K4905007"

Revision as of 11:02, 31 August 2023

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). This resulted in the sequence Z1-I[60]-A[100]-I[60]-Z2. More details about the components of this sequence can be found on their corresponding part pages. 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 (A[100]) and a hydrophobic region on each side of it (I[60]). 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.

figELP

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

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