Advantages of orange-labelled collagen and gelatine as substrates for rapid collagenase activity measurement.
A simple method for the determination of collagenase activity utilising dye-labelled substrates is proposed. It consists in labelling gelatine and bovine Achilles tendon collagen under nondenaturing conditions with the dye active orange GT. As verified with two different enzyme preparations, labelling did not dramatically change the susceptibility to collagenases. The kinetic parameters obtained for dye-labelled collagen and gelatine were compared to those obtained for the hydrolysis of native insoluble collagen (Mandl's method). The method offers the following advantages: it is rapid, reproducible, does not require special equipment and is more specific for collagenases than the widely used azocoll and Mandl methods.
Komsa-Penkova
R S
RS
Department of Chemistry and Biochemistry, Medical University, Pleven, Bulgaria.
Rashap
R K
RK
Yomtova
V M
VM
D000818:Animals; D001391:Azo Compounds / Q000378:metabolism; D002417:Cattle; D003094:Collagen / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism*; D004396:Coloring Agents / Q000378:metabolism*; D005780:Gelatin / Q000378:metabolism*; D007700:Kinetics; D012995:Solubility; D013379:Substrate Specificity
D001391:Azo Compounds; D004396:Coloring Agents; C028470:Azocoll; D005780:Gelatin; D003094:Collagen; D017364:Collagenases