Site-specific immobilization of protein layers on gold surfaces via orthogonal sortases

- Recombinant fluorescent proteins with the LPETG/A tags were site-specifically immobilized on gold surfaces modified with a short peptide tag using Sortase A.
- Layers of recombinant mCherry and GFP were immobilized on the gold surface layer-by-layer using Streptococcus pyogenes and Staphylococcus aureus Sortase A enzymes.
- Fluorescent microscopy, Western blotting, and atomic force microscopy were used to confirm sortase-mediated protein ligation and immobilization on the gold surface.