Engineering ultrastable protein scaffold for the controlled assembly of multifunctional nano-biomaterials
- Author(s): Lim, Samuel
- Advisor(s): Clark, Douglas S
- et al.
Self-assembling protein templates are of increasing interest in the field of nanoscale fabrication of biomaterials, where precise patterning of functional biomolecules, such as enzymes, is often desired. In particular, protein building blocks can be strategically chosen to exhibit desired functionality, while engineering their assembly allows for the controllable positioning of the subunits. The filamentous protein gamma-prefoldin (gPFD) from the hyperthermophilic archaeon Methanocaldococcus jannaschii is an excellent candidate for such a tunable scaffold. Its remarkable stability, unique modularity, and self-assembly into filaments with chaperone activity render it an ideal candidate for the bottom-up construction of novel protein nanostructures.
Our research aimed to construct functional protein biomaterials with precisely controlled nanostructures using gPFD as a building block. We engineered a versatile gPFD-based platform upon which scaffolded biocatalytic systems can be constructed in a customizable fashion. Furthermore, to gain precise positioning of functional molecules on our protein nanostructures, we developed multicomponent protein templates composed of distinct monomers that assemble in repeating orders; fusing different biomolecules to each subunit enabled periodic positioning of multiple functional features along the filament. Finally, we explored gPFD’s potential to form cross-linked network, and reported a gPFD-based functional hydrogel with tunable bulk properties. Ultimately, we expect the strategies developed in our lab to provide a gPFD-based biomolecular construction toolkit, which will enhance our ability to fabricate advanced multifunctional nanobiomaterials with novel chemical, catalytic, and structural properties.