Genetic Fusion of Thermoresponsive Polypeptides with UCST-type Behavior Mediates 1D Assembly of Coiled-Coil Bundlemers

Graphical Abstract: Available at: Computationally designed coiled coil-forming peptides were functionalized with thermoresponsive resilin-like polypeptides (RLPs) of various lengths and produced via biosynthetic methods in bacterial expression hosts. Interactions between RLPs upon cooling below their upper critical solution temperature (UCST) resulted in nanofibrillar assembly. Abstract Thermoresponsive resilin-like polypeptides (RLPs) of various lengths were genetically fused to two different computationally designed coiled coil-forming peptides with distinct thermal stability, to develop new strategies to assemble coiled coil peptides via temperature-triggered phase separation of the RLP units. Their successful production in bacterial expression hosts was verified via gel electrophoresis, mass spectrometry, and amino acid analysis. Circular dichroism (CD) spectroscopy, ultraviolet-visible (UV/Vis) turbidimetry, and dynamic light scattering (DLS) measurements confirmed the stability of the coiled coils and showed that the thermosensitive phase behavior of the RLPs was preserved in the genetically fused hybrid polypeptides. Cryogenic-transmission electron microscopy and coarse-grained modeling revealed that functionalizing the coiled coils with thermoresponsive RLPs leads to their thermally triggered noncovalent assembly into nanofibrillar assemblies.
This is the peer reviewed version of the following article: Patkar, S. S., Tang, Y., Bisram, A. M., Zhang, T., Saven, J. G., Pochan, D. J., Kiick, K. L., Angew. Chem. Int. Ed. 2023, e202301331, which has been published in final form at This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited. This article will be embargoed until 05/09/2024.
coiled coil peptides, peptides, phase transitions, resilin-like polypeptides, self-assembly
Patkar, S. S., Tang, Y., Bisram, A. M., Zhang, T., Saven, J. G., Pochan, D. J., Kiick, K. L., Angew. Chem. Int. Ed. 2023, e202301331.