Tryptophan-contained peptide-functional nanomaterials as general spectrofluorometric reagents for enzyme

By designing and coupling a functional peptide, Gly-Leu-Ala-Cys-Ser-Gly-Phe-Pro-Arg-Gly-Arg-Trp, which could be cleaved by thrombin at the site of Arg-Gly (R-G), to the surface of gold nanoparticles (Au-NPs), we propose a simple spectrofluorometry for thrombin (TRB) in this contribution. Experiments showed that the peptide coupled to the surface of Au-NPs in a Tris–HCl buffer at 37 °C could be cleaved, leaving the fluorescent fragment of Gly-Arg-Trp in the Au-NPs suspension. By centrifuging the suspension and measuring the fluorescence signals resulting from the Trp residue of Gly-Arg-Trp fragment in the supernatant, we found that the fluorescence intensity is proportional to thrombin concentrations in the range of 1–100 nM with the limit of the detection of 0.1 nM. Since there are a lot of enzymes that can hydrolyze peptide with special sequence, and novel nanomaterials that can bind with the tryptophan-contained peptide and understand centrifugation, this spectrofluorometric method is general and it is possible to develop a variety of detection method for target enzymes.