Beyond Dimerization: Harnessing Tetrameric Coiled‐Coils for Nanostructure Assembly

Versatile DNA and polypeptide-based structures have been designed based on complementary modules. However, polypeptides can also form higher oligomeric states. We investigated the introduction of tetrameric modules as a substitute for coiled-coil dimerization units used in previous modular nanostructures. Tetramerizing helical bundles can run in parallel or antiparallel orientation, expanding the number of topological solutions for modular nanostructures. Furthermore, this strategy facilitates the construction of nanostructures from two identical polypeptide chains. Importantly, tetrameric modules substantially stabilized protein nanostructures against air–water interface denaturation, enabling the determination of the first cryoelectron microscopy three-dimensional structure of a coiled-coil-based nanostructure, confirming the designed agreement of the modules forming a tetrahedral cage.