A continuous fluorimetric assay for clostridial collagenase and Pz-peptidase activity.
The peptide derivative N alpha-(2,4-dinitrophenyl)-L-prolyl-L-leucyl-glycyl-L-prolyl-L-tryptophanyl-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 (gamma ex. 283 nm, lambda em. 350 nm) following hydrolysis of the Leu2-Gly3 peptide bond. The value of Km for clostridial collagenase was 17 microM. The substrate for the first time makes possible continuous fluorimetric assays for Pz-peptidase and clostridial collagenase.
Barrett
A J
AJ
Biochemistry Department, Strangeways Research Laboratory, Cambridge, U.K.
Knight
C G
CG
Brown
M A
MA
Tisljar
U
U
D003013:Clostridium / Q000201:enzymology*; D010450:Endopeptidases / Q000032:analysis*; D005470:Fluorometry; D008666:Metalloendopeptidases; D003012:Microbial Collagenase / Q000032:analysis*
D010450:Endopeptidases; D008666:Metalloendopeptidases; C022510:thimet oligopeptidase; D003012:Microbial Collagenase