Open AccessHyperchromic effect of collagen induced by human collagenase
Author(s) -
LINDY Seppo,
SORSA Timo,
SUOMALAINEN Kimmo,
LAUHIO Anneli,
TURTO Heikki
Publication year - 1986
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1986.tb09539.x
Subject(s) - collagenase , hyperchromicity , collagen helix , enzyme , triple helix , chemistry , biophysics , denaturation (fissile materials) , cleavage (geology) , substrate (aquarium) , absorption (acoustics) , biochemistry , materials science , stereochemistry , nuclear chemistry , biology , dna , ecology , fracture (geology) , composite material
Loss of the highly ordered triple‐helix structure of native collagen on denaturation or enzymatic degradation involves a helix‐to‐coil transition, which can be seen as an increase at 227 nm in its ultraviolet difference absorption spectrum. We report here the successful use of this hyperchromic effect to quantify collagen in solution and to follow up the time‐course of collagen degradation catalyzed by collagenase. Using 14 C‐labelled collagen substrate we show the excellent correlation between enzyme‐induced increase in ultraviolet difference absorption and formation of specific cleavage products. The novel method was found to be suitable to characterize the enzymatic properties of human leukocyte collagenase. Activation of latent collagenase to the active enzyme could be followed continuously and an activation lag estimated.