The interaction of purified rabbit bone collagenase with purified rabbit bone metalloproteinase inhibitor

Abstract

1. Pure rabbit bone metalloproteinase inhibitor (TIMP) bound tightly to pure rabbit bone collagenase with an apparent Kd of 1.4 × 10(-10) M. 2. The molecular weight of the enzyme-inhibitor complex was found to be 54 000, but no enzyme activity could be recovered from the complex after treatment with either mercurials or proteinases. The complex thus differed from latent collagenase in terms of size, susceptibility to mercurials and behaviour on concanavalin A-Sepharose. 3. The interaction of the purified components was compared with that of crude collagenase and crude inhibitor in culture medium. Mercurial treatment partially reversed the inhibition in the crude system, but not when the purified components were used. 4. The significance of the results is discussed in relation to the extracellular control of the activity of collagenase.

Inactivation of tissue inhibitor of metalloproteinase‐1 by peroxynitrite

“…The activity of stromelysin-1 was measured using [14C]acetylated casein as substrate in 40 mM Tris-HCl (pH 7.6) with 12 mM CaC12 as described by Cawston et al [11]. Reaction mixtures were incubated for 20 h at 37°C and incorporated 0.93 ~tg/~tl stromelysin.…”

“…It inhibits active metalloproteinases, such as stromelysins, collagenases and gelatinases, by forming a tight one-to-one stoichiometric complex [11]. Thus, the balance between the activities of TIMPs and active metalloproteinases is probably a critical factor in the control of connective tissue remodelling in both health and disease, and a change of this balance in favour of active metalloproteinases may be an important determinant in the disease process.…”

The measurement of collagenase, tissue inhibitor of metalloproteinases (timp), and collagenase—timp complex in synovial fluids from patients with osteoarthritis and rheumatoid arthritis

“…TIMP is a glycoprotein of M, 28,000 containing 12 cysteine residues which form 6 disulfide bonds (8). It forms 1 : 1 noncovalent complexes with all of the active MMPs, with a Ki of lo-'' M for collagenase (9). TIMP is responsible for preventing uncontrolled breakdown of connective tissues; agents that promote TIMP secretion, when added to resorbing cartilage, reduce the release of glycosaminoglycan fragments from the cartilage (10).…”

Proteolysis in human breast and colorectal cancer

“…Four human TIMPs have been identified to date (TIMPs 1-4) which have closely related structures and inhibitory properties. TIMPs bind non-covalently to active MMPs forming essentially irreversible complexes inhibiting MMP activity (Cawston et al, 1983). Certain TIMPs also bind to latent forms of MMPs, e.g.…”