A Self-Driving and Self-Reporting Petal-Like Au–Cu ₂ O Metalloenzyme for Probing H ₂ S-Mediated Cuproptosis

Hydrogen sulfide (H₂S), highly enriched in colorectal tumors, acts as an upstream regulator of copper homeostasis and cuproptosis. However, most existing cuproptosis nanotherapeutics focus on downstream copper overload while lacking the ability to resolve the dynamic H₂S-mediated regulation that governs copper speciation and redox stress. Here, we develop a self-driving and self-reporting petal-like Au-Cu₂O metalloenzyme that enables real-time interrogation of H₂S-mediated cuproptosis. Surfactant-directed anisotropic growth yields an interface-rich architecture with exposed Au-Cu₂O junctions, generating abundant plasmonic hotspots and redox-active sites for synergistic SERS enhancement and photoenhanced peroxidase-like catalysis. The nanocomposite drives sustained Cu⁺/Cu²⁺ cycling, glutathione depletion, and reactive oxygen species generation, leading to mitochondrial dysfunction and lipid peroxidation. Using activity-based SERS monitoring in living cells, we reveal that H₂S exerts a dual regulatory role by transiently buffering oxidative stress while promoting intracellular copper retention through copper-sulfide complexation, thereby amplifying downstream cuproptosis execution. By correlating H₂S upregulation with copper retention and cuproptosis markers in colorectal cancer models, this work establishes a foundation for precision intervention against H₂S-altered malignancies.