Latest revision as of 12:31, 2 October 2023

Elastin-like Polypeptide (VPGIG)[60]

Information

Elastin-like polypeptides (ELPs) are protein polymers derived from human tropoelastin. One of their key features is that they exhibit a phase separation that is often reversible whereby samples remain soluble below a transition temperature (T_t) but form coacervates above T_t. They have many possible applications in purification, sensing, activation, and nano assembly. Furthermore, they are non-immunogenic, substrates for proteolytic biodegradation, and can be decorated with pharmacologically active peptides, proteins, and small molecules. Recombinant synthesis additionally allows precise control over ELP architecture and molecular weight, resulting in protein polymers with uniform physicochemical properties suited to the design of multifunctional biologics. As such, ELPs have been employed for various uses including as anti-cancer agents, ocular drug delivery vehicles, and protein trafficking modulators^[2].

The general structure of polymeric ELPs is (VPGXG)_n, where the monomeric unit is Val-Pro-Gly-X-Gly, and the "X" denotes a variable amino acid that can have consequences on the general properties of the ELP, such as the T_t. Specifically, the hydrophilicity or hydrophobicity and the presence or absence of a charge on the guest residue play a great role in determining the T_t. Also, the solubilization of the guest residue can affect the Tt. The "n" denotes the number of monomeric units that comprise the polymer^[1]. In general, these polymers are linear below the Tt, but aggregate into spherical clumps above the Tt^[3].The repeating sequence of this part is (VPGIG)_[60], also referred to as I_[60], which creates a hydrophobic part polymer.

The TU-Eindhoven 2023 team used this part in a composite part where it was fuzed to mneongreen ( BBa_K1761003) to form the composite part BBa_K4905016.

The sequence was designed and codon optimized for E.coli BL21 cells. The DNA was transformed and multiplied in TOP10 competent cells.

References

[1] Christensen, T., Hassouneh, W., Trabbic-Carlson, K., & Chilkoti, A. (2023). Predicting Transition Temperatures of Elastin-Like Polypeptide Fusion Proteins. https://doi.org/10.1021/bm400167h

[2] 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

[3] Hassouneh, W., Christensen, T., & Chilkoti, A. (2010). Elastin-Like Polypeptides as a Purification Tag for Recombinant Proteins. Current Protocols in Protein Science, 61(1), 6.11.1-6.11.16. https://doi.org/10.1002/0471140864.PS0611S61