Preparation of a novel metallothionein-AuNP composite material by genetic modification and Au[sbnd]S covalent combination.

Metallothionein (MTs) can be used in the prevention and treatment of tumors and diabetes due to its antioxidant properties. However, it is necessary to solve its non-transmembrane properties and further improve its antioxidant activity, increase its fluorescence visualization and enhance its stability to meet practical applications in the biomedical field. Here, we report the preparation of a novel metallothionein-AuNP composite material with high transmembrane ability, fluorescence visualization, antioxidant activity, and stability by genetic modification (introducing transduction peptide TAT, fluorescence tag GFP and increasing sulfydryl groups) and immobilization technology (covalently bonding with AuNPs). The transmembrane activity of modified proteins was verified by immunofluorescence. Increasing the sulfhydryl content within a certain range can enhance the antioxidant activity of the protein. In addition, GFP were used to further simplify the imaging of the metallothionein-AuNP composite in cells. XPS results indicated that AuNPs can immobilize metallothionein through Au S covalent bonds. TGA characterization and degradation experiments showed that thermal and degradation stability of the immobilized material was significantly improved. This work provides new ideas to construct metallothionein composites with high transmembrane ability, antioxidant activity, fluorescence visualization and stability to meet novel applications in the biomedical field. [Display omitted] • TAT- Sh MT3 was obtained by long primer PCR. • TAT- Sh MT3 has about 30 % increase in antioxidant capacity compared to TAT- Sh MT. • TAT- Sh MT3-GFP was obtained by SOE-PCR. • TAT- Sh MT3-GFP + AuNPs was obtained Au S covalent bond. • TAT-ShMT3-GFP + AuNPs has higher stability compared to TAT- Sh MT3-GFP. [ABSTRACT FROM AUTHOR]