Engineering protein polymers of ultrahigh molecular weight via supramolecular polymerization: towards mimicking the giant muscle protein titin

Abstract

Utilizing protein fragment reconstitution of a small protein GB1, we developed an efficient, supramolecular polymerization strategy to engineer protein polymers with ultrahigh molecular weight that mimic the giant muscle protein titin.

Reaction Rate Governs the Viscoelasticity and Nanostructure of Folded Protein Hydrogels

“…So-called folded protein hydrogels are capable of reproducing a variety of functional properties such as mimicking the nonlinear elastic response of muscles, − maintaining their mechanical integrity even after large deformations , or after exposure to chemical reduction, and to be mechanically responsive to chemical stimuli . The design of these hydrogels depends on translating the precise molecular scale mechanical response of the individual proteins into bulk mechanical properties through recombinant design. , While such approaches have been shown to be both precise and effective in creating folded proteins with desired mechanical properties, they have a number of limitations. Both the recombinant design and the subsequent expression and purification of the protein typically require significant optimization.…”

Non-Covalently Associated Streptavidin Multi-Arm Nanohubs Exhibit Mechanical and Thermal Stability in Protein-Network Materials

“…This customizable platform design can be engineered at the genetic level to incorporate target proteins with key mechanical, or biochemical properties in protein-network strands, or investigate alternative stimuli-induced chemical cross-linkers, such as GB1, 26 to further improve secondary crosslinking specificity. Moreover, biotinylated molecules can be orthogonally coordinated with the SAv-based protein-network system by using the strong protein-ligand interaction of SAv-biotin for diverse biomedical applications, such as advanced cell signaling, drug delivery, biosensing, and imaging.…”

“…The systematic characterization of fourarm SAv-based hierarchical nanoassembly using a variety of second-level molecular selfassembly or specific chemical crosslinking methods discovered that stimuli-induced chemical cross-linkers, such as dityrosine photo-crosslinking, are optimal for developing protein-network materials with enhanced specificity, stability, and network homogeneity. This customizable platform design can be engineered at the genetic level to incorporate target proteins with key mechanical, or biochemical properties in protein-network strands, or investigate alternative stimuli-induced chemical cross-linkers, such as GB1, 26 to further improve secondary crosslinking specificity. Moreover, biotinylated molecules can be orthogonally coordinated with the SAv-based protein-network system by using the strong protein-ligand interaction of SAv-biotin for diverse biomedical applications, such as advanced cell signaling, drug delivery, biosensing, and imaging.…”

“…22 Secondary or tertiary structures of proteins that self-recognize and non-covalently selfassociate each other, constituting quaternary or oligomeric structures that have been used as biomolecular cross-linkers. By genetically fusing the cross-linkers to proteins of interest using recombinant DNA technology, biosynthesized artificial proteins can construct predictable macromolecular structures, [23][24][25][26] such as multi-arm star-like precursors similar to tetra-PEG crosslinkers (𝑓 ≥ 3). 23 Furthermore, this approach enables the fabrication of protein-network materials with cross-linkers containing tailored binding affinity, modulating the intrinsic nature of protein associations and strands with customized length and mechanics.…”

There Is Plenty of Room in The Folded Globular Proteins: Tandem Modular Elastomeric Proteins Offer New Opportunities in Engineering Protein‐Based Biomaterials

“…[ 83 ] Similarly, increasing the molecular weight of SAE proteins will likely lead to improvements in the mechanical properties of SAE protein hydrogels. [ 84 ] For this, postexpression polymerization strategy, which further polymerizes recombinantly expressed SAE proteins, as well as new expression systems, which likely involves bacterial engineering, will be desirable.…”