Diversification of Protein Cage Structure Using Circularly Permuted Subunits

• Published in: Journal of the American Chemical Society Vol. 140; no. 2; pp. 558 - 561
• Main Authors: Azuma, Yusuke, Herger, Michael, Hilvert, Donald
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 17-01-2018
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/jacs.7b10513

Self-assembling protein cages are useful as nanoscale molecular containers for diverse applications in biotechnology and medicine. To expand the utility of such systems, there is considerable interest in customizing the structures of natural cage-forming proteins and designing new ones. Here we report that a circularly permuted variant of lumazine synthase, a cage-forming enzyme from Aquifex aeolicus (AaLS) affords versatile building blocks for the construction of nanocompartments that can be easily produced, tailored, and diversified. The topologically altered protein, cpAaLS, self-assembles into spherical and tubular cage structures with morphologies that can be controlled by the length of the linker connecting the native termini. Moreover, cpAaLS proteins integrate into wild-type and other engineered AaLS assemblies by coproduction in Escherichia coli to form patchwork cages. This coassembly strategy enables encapsulation of guest proteins in the lumen, modification of the exterior through genetic fusion, and tuning of the size and electrostatics of the compartments. This addition to the family of AaLS cages broadens the scope of this system for further applications and highlights the utility of circular permutation as a potentially general strategy for tailoring the properties of cage-forming proteins.