Zn-doped Cu₂S quantum dots as new high-efficiency inhibitors against human insulin fibrillation based on specific electrostatic interaction with oligomers

Inhibition of protein fibrillation process with nanomaterials is a promising strategy to combat neurodegenerative diseases. Copper-based nanomaterials have been seldom utilized in fibrillation inhibiting research due to Copper ions are generally considered as accelerators of fibrosis. Here, we proposed ultra-small Zn doped Cu₂S (Zn:Cu₂S) QDs as inhibitors of human insulin (HI) fibrosis. ThT, DLS, CD and TEM confirm that Zn:Cu₂S QDs effectively inhibited insulin fibrosis in a dose-dependent manner with lag phase time extended (beyond 13-time by Zn:Cu₂S QDs of 1 mg·mL^-1), final fibril formation and the conversion from α-helix to β-sheet reduced. Additionally, thermodynamics analyzed results reveal that the HI fluorescence quenching process is static quenching dominated, and the Zn:Cu₂S QDs inhibit HI fibrosis mainly through specific electrostatic interaction with oligomers. The positively charged amino acid residues of oligomers bind to the negatively charged Zn:Cu₂S QDs, which prevents the self-assembly of the oligomers from growing into mature fibers to enhance the stability of the protein. Unlike free Copper ions, the as-prepared QDs show an excellent inhibition in HI fibrillation, breaking through the bottleneck of copper-based materials in inhibiting protein fibrosis and providing a potential strategy to inhibit protein fibrosis in-situ by biosynthesizing copper-based fibrosis inhibitors.