De novo design of modular protein hydrogels with programmable intra- and extracellular viscoelasticity

• 成果类型: Article
• 署名作者: Mout, Rubul; Bretherton, Ross C.; Decarreau, Justin; Lee, Sangmin; Gregorio, Nicole; Edman, Natasha I.; Ahlrichs, Maggie; Hsia, Yang; Sahtoe, Danny D.; Ueda, George; Sharmal, Alee; Schulman, Rebecca; Deforest, Cole A.; Bakera, David
• 署名单位: University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; Harvard University; Harvard University Medical Affiliates; Boston Children's Hospital; Harvard Medical School; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; University of Washington; University of Washington Seattle; Howard Hughes Medical Institute; Northeastern University; Johns Hopkins University; Johns Hopkins University
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-12631
• DOI: 10.1073/pnas.2309457121
• 发表日期: 2024-02-01
• 关键词: neutralizing antibody-responses; computational design; accurate design; software suite; stem-cell; simulation; DYNAMICS; bond

摘要

Relating the macroscopic properties of protein - based materials to their underlying component microstructure is an outstanding challenge. Here, we exploit computational design to specify the size, flexibility, and valency of de novo protein building blocks, as well as the interaction dynamics between them, to investigate how molecular parameters govern the macroscopic viscoelasticity of the resultant protein hydrogels. We construct gel systems from pairs of symmetric protein homo- oligomers, each comprising 2, 5, 24, or 120 individual protein components, that are crosslinked either physically or covalently into idealized step- growth biopolymer networks. Through rheological assessment, we find that the covalent linkage of multifunctional precursors yields hydrogels whose viscoelasticity depends on the crosslink length between the constituent building blocks. In contrast, reversibly crosslinking the homo- oligomeric components with a computationally designed heterodimer results in viscoelastic biomaterials exhibiting fluid - like properties under rest and low shear, but solid - like behavior at higher frequencies. Exploiting the unique genetic encodability of these materials, we demonstrate the assembly of protein networks within living mammalian cells and show via fluorescence recovery after photobleaching (FRAP) that mechanical properties can be tuned intracellularly in a manner similar to formulations formed extracellularly. We anticipate that the ability to modularly construct and systematically program the viscoelastic properties of designer protein - based materials could have broad utility in biomedicine, with applications in tissue engineering, therapeutic delivery, and synthetic biology.