AbstractHerein, 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 NP.