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

Simple item page
Author(s)Patkar, Sai S.
Author(s)Tang, Yao
Author(s)Bisram, Arriana M.
Author(s)Zhang, Tianren
Author(s)Saven, Jeffery G.
Author(s)Pochan, Darrin J.
Author(s)Kiick, Kristi L.
Date Accessioned2023-05-26T19:39:51Z
Date Available2023-05-26T19:39:51Z
Publication Date2023-05-09
DescriptionThis 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 https://doi.org/10.1002/anie.202301331. 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.
AbstractGraphical Abstract: Available at: https://doi.org/10.1002/anie.202301331 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.
SponsorThis research was primarily supported by National Science Foundation through the University of Delaware Materials Research Science and Engineering Center, DMR-2011824. Partial support for the work was from DMR-1609544; DMR-2004796; DMR-0239744; DMR-2004890; and the National Institutes of Health (P20 RR017716; RO1 DC011377). Additional support for the work was from cooperative agreement #370NANB17H302 from NIST, U.S. Department of Commerce. The authors also acknowledge the financial support from the Department of Energy, Office of Basic Energy Sciences, Biomolecular Materials Program under grant nos. DESC0019355 and DE-SC0019282.The contents of this manuscript are solely the responsibility of the authors and do not necessarily reflect the views of the funding agencies. The authors would like to thank Haofu Huang for some preliminary cast-film TEM data demonstrating potential nanofibril formation, Joshua Meisenhelter from the Christopher Kloxin Research Group for providing SPPS control for BFPa and Matthew Langenstein from the Pochan group for providing SPPS controls (BFPb and RLP1). This work used Stampede 2 through allocation TG-CHE110041from the Advanced Cyberinfrastructure Coordi-nation Ecosystem: Services & Support (ACCESS) and XSEDE programs, which is supported by National Science Foundation grants #1548562, #2138259, #2138286, #2138307, #2137603, and #2138296.
CitationPatkar, S. S., Tang, Y., Bisram, A. M., Zhang, T., Saven, J. G., Pochan, D. J., Kiick, K. L., Angew. Chem. Int. Ed. 2023, e202301331.
ISSN1521-3773
URLhttps://udspace.udel.edu/handle/19716/32801
Languageen_US
PublisherAngewandte Chemie International Edition
Keywordscoiled coil peptides
Keywordspeptides
Keywordsphase transitions
Keywordsresilin-like polypeptides
Keywordsself-assembly
TitleGenetic Fusion of Thermoresponsive Polypeptides with UCST-type Behavior Mediates 1D Assembly of Coiled-Coil Bundlemers
TypeArticle
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
Genetic Fusion of Thermoresponsive Polypeptides.pdf
Size:
2.68 MB
Format:
Adobe Portable Document Format
Description:
Main article
Download
License bundle
Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
2.22 KB
Format:
Item-specific license agreed upon to submission
Description:
Download
Collections
Open Access Publications