A continuous fluorimetric assay for clostridial collagenase and Pz-peptidase activity. 1989

A J Barrett, and C G Knight, and M A Brown, and U Tisljar
Biochemistry Department, Strangeways Research Laboratory, Cambridge, U.K.

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.

UI MeSH Term Description Entries
D008666 Metalloendopeptidases ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism. Metallo-Endoproteinases,Metalloendopeptidase
D010450 Endopeptidases A subclass of PEPTIDE HYDROLASES that catalyze the internal cleavage of PEPTIDES or PROTEINS. Endopeptidase,Peptide Peptidohydrolases
D003012 Microbial Collagenase A metalloproteinase which degrades helical regions of native collagen to small fragments. Preferred cleavage is -Gly in the sequence -Pro-Xaa-Gly-Pro-. Six forms (or 2 classes) have been isolated from Clostridium histolyticum that are immunologically cross-reactive but possess different sequences and different specificities. Other variants have been isolated from Bacillus cereus, Empedobacter collagenolyticum, Pseudomonas marinoglutinosa, and species of Vibrio and Streptomyces. EC 3.4.24.3. Clostridiopeptidase A,Clostridium histolyticum Collagenase,Collagenase, Microbial,Collagenase Clostridium histolyticum,Collagenase-Like Peptidase,Collalysine,Nucleolysin,Clostridium histolyticum, Collagenase,Collagenase Like Peptidase,Collagenase, Clostridium histolyticum,Peptidase, Collagenase-Like,histolyticum, Collagenase Clostridium
D003013 Clostridium A genus of motile or nonmotile gram-positive bacteria of the family Clostridiaceae. Many species have been identified with some being pathogenic. They occur in water, soil, and in the intestinal tract of humans and lower animals.
D005470 Fluorometry An analytical method for detecting and measuring FLUORESCENCE in compounds or targets such as cells, proteins, or nucleotides, or targets previously labeled with FLUORESCENCE AGENTS. Fluorimetry,Fluorometric Analysis,Analysis, Fluorometric

Related Publications

A J Barrett, and C G Knight, and M A Brown, and U Tisljar
March 1990, The Biochemical journal,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
November 1985, Journal of chromatography,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
November 1979, Analytical biochemistry,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
November 1977, Wiener klinische Wochenschrift,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
September 1985, Journal of reproduction and fertility,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
October 1998, Analytical biochemistry,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
January 1980, Analytical biochemistry,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
May 1981, Analytical biochemistry,
A J Barrett, and C G Knight, and M A Brown, and U Tisljar
May 1990, Biological chemistry Hoppe-Seyler,
Copied contents to your clipboard!