Studies on collagen. II. Properties of purified collagenase and its inhibition.

In previous papers from these laboratories (1, 2) methods for the purification of the collagenase of Clostridiurm histolyticum were presented, and a calcium ion requirement for the binding of the enzyme to collagen and for the overall enzymatic action was reported. In the present, communication an additional method of purification, which makes possible the large-scale preparation of the enzyme, is described. The sedimentation and diffusion constants and molecular weight of the purified enzyme are also reported. The nature of the end products resulting from the action of collagenase on the citrate-extractable collagens of carp swim bladder and calfskin, as well as on the gelatins derived from these, is considered. We shall also describe the action of the enzyme on collagen in the presence of its derived gelatin; the failure of the enzyme to act on a number of synthetic peptides, amino acid polymers, and esters; and the inhibition of purified collagenase by sulfhydryl-containing substances and by certain metal-sequestering agents.