Hydrogels have a broad range of applications in biotechnology because of their biocompatibility, tunable viscoelasticity, and versatile methods of fabrication. Peptide motifs or protein domains are often incorporated into the hydrogel network in order to provide functionalities such as catalytic activity or stimuli-responsiveness. However, integrating combinations of functional proteins into the hydrogel in stable, customizable, and tunable fashion remains challenging. We aim to meet all three criteria by using self-assembling, crosslinkable protein scaffolds as the hydrogel backbone. The filamentous protein G-prefoldin (G-PFD) from the hyperthermophilic archaeon Methanocaldococcus jannaschii is an excellent candidate for such a modular protein template. Its remarkable stability, unique structure, and self-assembly into filaments render it an ideal candidate for the bottom-up construction of functional supramolecular biomaterials. We have engineered a terminal cysteine-containing variant of G-PFD, and shown that filaments composed of this subunit can be crosslinked to form hydrogels. Properties of the resulting hydrogel were highly tunable through varying the monomer concentration as well as the crosslink density. Furthermore, we have demonstrated that combinations of multiple proteins can be attached along the G-PFD filaments using the SpyCatcher-SpyTag chemistry. We aim to expand this approach to generate enzymatic hydrogels capable of supporting cascade reactions. Ultimately, our strategy will enable the design of smart hydrogel biomaterials for complex applications that require multifunctionalities, such as biosensors or bioelectronic devices.

Modular design and functionalization of enzymatic hydrogels using a self-assembling protein building block

253rd National Meeting (2017)

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