Herein, we report the synthesis and isolation of a shape-persistent Janus protein nanoparticle derived from biomolecular machine chaperonin GroEL (A GroEL B) and its application to DNA-mediated ternary supramolecular copolymerization. For synthesizing A GroEL B with two different DNA strands A and B at its opposite apical domains, we utilized the unique biological property of GroEL, i.e., "Mg 2+ /ATPmediated ring exchange" between A GroEL A and B GroEL B with their hollow cylindrical doubledecker architectures. This exchange event was reported more than 24 years ago but has never been utilized for molecular engineering of GroEL. We leveraged DNA nanotechnology to purely isolate Janus A GroEL B and succeeded in its precision ternary supramolecular copolymerization with two DNA comonomers A** and B* that are partially complementary to A and B in A GroEL B , respectively, and programmed to self-dimerize on the other side. Transmission electron microscopy allowed us to confirm the formation of an expected dual-periodic copolymer sequence-(B*/B GroEL A/A**/A**/A GroEL B/B*)-in the form of a laterally connected lamellar assembly, rather than a single-chain copolymer.

Herein, we report an ATP‐responsive nanoparticle (GroELNP) whose surface is fully covered with the biomolecular machine “chaperonin protein GroEL”. GroELNP was synthesized by DNA hybridization between a gold NP with DNA strands on its surface and GroEL carrying complementary DNA strands at its apical domains. The unique structure of GroELNP was visualized by transmission electron microscopy including under cryogenic conditions. The immobilized GroEL units retain their machine‐like function and enable GroELNP to capture denatured green fluorescent protein and release it in response to ATP. Interestingly, the ATPase activity of GroELNP per GroEL was 4.8 and 4.0 times greater than those of precursor cysGroEL and its DNA‐functionalized analogue, respectively. Finally, we confirmed that GroELNP could be iteratively extended to double‐layered (GroEL)2 ${{^{({\rm GroEL}){_{2}}}}}$ NP.

Herein, we report an ATP-responsive nanoparticle ( GroEL NP) whose surface is fully covered with the biomolecular machine "chaperonin protein GroEL". GroEL NP was synthesized by DNA hybridization between a gold NP with DNA strands on its surface and GroEL carrying complementary DNA strands at its apical domains. The unique structure of GroEL NP was visualized Angewandte Chemie