A continuous fluorimetric assay for clostridial collagenase and Pz-peptidase activity

Abstract

The peptide derivative N(α)-(2,4-dinitrophenyl)-L-prolyl-L-leucyl-glycyl-L-prolyl-L-tryptop anyl-D-lysine (Dnp-Pro-Leu-Gly-Pro-Trp-D-Lys) has been found to be a convenient substrate for the assay of clostridial collagenase and Pz-peptidase. The substrate shows a 25-fold enhancement of fluorescence (λ(ex.) 283 nm, λ(em.) 350 nm) following hydrolysis of the Leu2-Glya3 peptide bond. The value of K(m) for clostridial collagenase was 17 μM. The substrate for the first time makes possible continuous fluorimetric assays for Pz-peptidase and clostridial collagenase.