Protein Cages

Self-assembling protein cages are widespread in biology. By providing a protective shell for cargo molecules, these nanostructures serve as multipurpose containers for the transport of viral genomes, mineralization, protein folding and degradation, and catalysis of short metabolic sequences. As exemplified by their broad functionality in nature, these protein cages provide valuable platforms for development of bionanotechnology. We have tailored and repurposed such proteinaceous compartments as reaction chambers for catalysis, vehicles for delivery/display, and building blocks for novel nanoarchitectures. For instance, a bacterial protein cage has been recently engineered and evolved to form primordial mimicries of carboxysome, proteasome, and virus-like structures that encapsulate own genetic information. Such bottom-up approaches to construct biomimetic compartments afford useful insights into principles underlying molecular mechanism of natural counterparts, and will pave the way for prospective applications in biology and medicine.

Selected Publications

Edwardson, T. G. W., Mori, T., and Hilvert, D. Rational Engineering of a Designed Protein Cage for siRNA Delivery. J. Am. Chem. Soc., 140(33):10439-10442.

Terasaka, N., Azuma, Y., and Hilvert, D. Laboratory evolution of virus-like nucleocapsids from nonviral protein cages. Proc. Natl. Acad. Sci. U.S.A, 115(21), 5432–5437 (2018).

Azuma, Y., Edwardson, T. G. W., and Hilvert, D. Tailoring lumazine synthase assemblies for bionanotechnology. Chemical Society Reviews, 47(10), 3543–3557 (2018).

Azuma, Y., Bader, D. L. V., and Hilvert, D. Substrate Sorting by a Supercharged Nanoreactor. J. Am. Chem. Soc., 140(3), 860–863 (2018).

Azuma, Y., Herger, M., and Hilvert, D. Diversification of Protein Cage Structure Using Circularly Permuted Subunits. J. Am. Chem. Soc., 140(2), 558–561 (2018).

Azuma, Y., Edwardson, T. G. W., Terasaka, N., and Hilvert, D. Modular Protein Cages for Size-Selective RNA Packaging in Vivo. J. Am. Chem. Soc., 140(2), 566–569 (2018).

Azuma, Y., Hilvert, D. Enzyme Encapsulation in an Engineered Lumazine Synthase Protein Cage. Methods Mol. Biol, 1798(Chapter 4), 39–55 (2018).

Tetter, S., and Hilvert, D. Enzyme Encapsulation by a Ferritin Cage. Angew. Chem. Int. Ed., 56(47), 14933–1493. (2017).

Sasaki, E., Böhringer, D., van de Waterbeemd, M., Leibundgut, M., Zschoche, R., Heck, A. J. R., et al. Structure and assembly of scalable porous protein cages. Nat. Comm., 8, 1–10 (2017).

Azuma, Y., Zschoche, R., and Hilvert, D. The C-terminal peptide of Aquifex aeolicusriboflavin synthase directs encapsulation of native and foreign guests by a cage-forming lumazine synthase. J. Biol. Chem., 292(25), 10321–10327 (2017).

Frey, R., Mantri, S., Rocca, M., and Hilvert, D. Bottom-up Construction of a Primordial Carboxysome Mimic. J. Am. Chem. Soc., 138(32), 10072–10075 (2016).

Sasaki, E., and Hilvert, D. Self-Assembly of Proteinaceous Multishell Structures Mediated by a Supercharged Protein. J. Phys. Chem. B, 120(26), 6089–6095 (2016).

Azuma, Y., Zschoche, R., Tinzl, M., and Hilvert, D. Quantitative Packaging of Active Enzymes into a Protein Cage. Angew. Chem. Int. Ed., 55(4), 1531–1534 (2016).

Frey, R., Hayashi, T., and Hilvert, D. Enzyme-mediated polymerization inside engineered protein cages. Chemical Communications, 52(68), 10423–10426 (2016).

Zschoche, R., and Hilvert, D. Diffusion-Limited Cargo Loading of an Engineered Protein Container. J. Am. Chem. Soc., 137(51), 16121–16132 (2015).

Beck, T., Tetter, S., Künzle, M., and Hilvert, D. Construction of Matryoshka-type structures from supercharged protein nanocages. Angew. Chem. Int. Ed., 54(3), 937–940 (2015).

Wörsdörfer, B., Pianowski, Z., and Hilvert, D. Efficient in vitro encapsulation of protein cargo by an engineered protein container. J. Am. Chem. Soc., 134(2), 909–911 (2012).

Wörsdörfer, B., Woycechowsky, K. J., and Hilvert, D. Directed evolution of a protein container. Science, 331(6017), 589–592 (2011).

Seebeck, F. P., Woycechowsky, K. J., Zhuang, W., Rabe, J. P., and Hilvert, D. A simple tagging system for protein encapsulation. J. Am. Chem. Soc., 128(14), 4516–4517 (2006).