Engineering Protein Hydrogels Using SpyCatcher-SpyTag Chemistry

Abstract

Constructing hydrogels from engineered proteins has attracted significant attention within the material sciences, owing to their myriad potential applications in biomedical engineering. Developing efficient methods to cross-link tailored protein building blocks into hydrogels with desirable mechanical, physical, and functional properties is of paramount importance. By making use of the recently developed SpyCatcher-SpyTag chemistry, we successfully engineered protein hydrogels on the basis of engineered tandem modular elastomeric proteins. Our resultant protein hydrogels are soft but stable, and show excellent biocompatibility. As the first step, we tested the use of these hydrogels as a drug carrier, as well as in encapsulating human lung fibroblast cells. Our results demonstrate the robustness of the SpyCatcher-SpyTag chemistry, even when the SpyTag (or SpyCatcher) is flanked by folded globular domains. These results demonstrate that SpyCatcher-SpyTag chemistry can be used to engineer protein hydrogels from tandem modular elastomeric proteins that can find applications in tissue engineering, in fundamental mechano-biological studies, and as a controlled drug release vehicle.

Engineering Potato Virus X Particles for a Covalent Protein Based Attachment of Enzymes

“…[54] This system has been used for many different approaches, including the preparation of catalytic biofilms, immunization, hydrogel engineering, and the cyclization of proteins. [55][56][57][58][59][60][61][62] The small size of the ST and its specific and irreversible isopeptide bond formation with SC in a wide range of buffers and over a wide temperature range makes this complementing protein pair ideal for the specific attachment of target peptides to the surface of viral nanoparticles. [54] Furthermore, the reaction may be sequentially coupled with individual target peptides fused to SC, allowing a variety of proteins to be displayed.…”

Bottom-up approaches in synthetic biology and biomaterials for tissue engineering applications

“…Altogether, the SpyCatcher-SpyTag is a modular system can be used for a variety of applications to produce novel protein architectures, including joining functionalized polypeptide strands to proteins (Fig. 4C) at specific times during cell culture [230], provide an environment to improve viability of cells [232], enzyme stability [233], and vaccine optimization [234]. …”

B 12 -dependent photoresponsive protein hydrogels for controlled stem cell/protein release

“…Assembling genetically engineered proteins into molecular networks represents an alternative strategy to make hydrogels with well-controlled properties (16)(17)(18)(19). Although natural evolution has led to numerous functional protein domains that can sense and respond to a variety of environmental stimuli, such as light, oxidative stress, pH, small molecules, metal ions, etc., such ecological diversity has yet to be fully tapped to develop responsive biomaterials with dynamically tunable properties.…”

“…Because of its high efficiency and modularity, this chemistry has led to a number of applications, including control of biomacromolecular topology, synthesis of bioactive and "living" materials, and biomolecular imaging (16,18,(27)(28)(29)(30)(31)(32)(33)(34)(35)(36). It has proven to be a powerful method for constructing complex biomolecular architectures both in vitro and in vivo.…”