MMP-13 (collagenase 3) is a newly discovered matrix metalloproteinase found in various tissues such as malignant tumors, osteoarthritic cartilage, rheumatoid synovium and wounds. MMP-13 production in chondroctyes and synoviocytes is upregulated by stimulation with inflammatory mediators such as IL-1, TNF and retinoic acid (reference 1). MMP-13 has been shown to degrade type I and II collagen and

the degradation of type II collagen occurs approximately ten times faster than that of type I collagen (reference 2). The typical 3/4 and 1/4 fragements of collagen are produced as with MMP-1 (collagenase 1). However, MMP-13 also generates a second cleavage of type II collagen that removes three amino acids from the amino terminus of the 1/4 fragement. In addition, MMP-13 degrades aggrecan, the major

proteoglycan of cartilage (reference 3).

Inhibitors of MMPs may be useful therapeutics to prevent metastasis of certain cancer cells and tissue damage in inflammatory diseases. This MMP-13 inhibitor assay kit is ideal for assessing inhibitors of MMP-13 and related enzymes. MMP-13 inhibitors can be easily screened and evaluated by simply adding potential inhibitory compounds to the activated recombinant human MMP-13 (rhMMP-13), see MMP-13 inhibitor assay sheet, page 5. This kit contains a truncated, rhMMP-13, which cleaves a fluorogenic peptide substrate included in this kit, but is not capable of digesting native collagen molecules. This kit contains all necessary reagents to perform 100 reactions.

Furthermore, this kit is useful for assaying total collagenase-like proteinase activity in tissue fluids or extracts and cell culture supernatants. Since the fluorogenic peptide substrate used in this kit does not hold a triple helical structure, this substrate is not specific to collagenase and

might be susceptible to non-collagenolytic proteinases. However, it is important to know the total activity of collagenase-like proteinases in specimens before proceeding with further experiments.