Demonstration of tissue collagenase activity in vivo and its relationship to inflammation severity in human gingiva

Abstract

Overall CM, Wiebkin O W, and Thonard JC: Demonstration of tissue collagenase activity in vivo and its relationship to inflammation severity in human gingiva. Journal of Periodontal Research; 1987: 22: 81-88. To directly demonstrate both the presence and in vivo activity of tissue collagenase (EC 3.4.24.7) during gingival inflammation, tissue extracts from 54 specimens of variously inflamed human gingiva were analyzed individually for: a) collagenasespecific collagen degradation products, and b) collagen-bound collagenase. The TC"^ collagen degradation product, identified using SDS-PAGE, was shown in 13/19 (68.4%) of insoluble tissue-residue fractions extracted from moderate-toseverely inflamed gingiva, but in only 2/21 (9.6%) slight-to-mildly inflamed gingival specimens, suggesting differences in the in vivo collagenase activity between the two groups. Using in vitro collagenase assays, gingival collagenase (as bound to insoluble collagen) was demonstrated in 92.8% of the moderate-toseverely inflamed gingival specimens and in 50% of the slight-to-mildly inflamed gingival specimens. A relationship was established between the active and latent forms of the enzyme and the degree of inflammation. Active enzyme was present in 78% of the moderate-to-severely inflamed specimens and in 14% of the slightto-mildly inflamed gingiva. In contrast, latent collagenase was predominant in the slight-to-mildly inflamed group (86% of coliagenase-positive samples) compared with 46% of coliagenase-positive moderate-to-severely inflamed gingival samples. The collagen-bound gingival collagenase was inhibited by metal ion chelators, sulphydryl reagents and 10% FBS, but not by serine-nor thiol-proteinase inhibitors and is therefore a neutral metalloproteinase.

Membrane protease proteomics: Isotope-coded affinity tag MS identification of undescribed MT1–matrix metalloproteinase substrates

“…Hashimoto et al (41) also recently described CTGF cleavage by soluble MMPs 1, 3, 7, and 13 at Met-194-Ile-195 within the CTGF linker, with a minor cleavage site at Ala-181-Tyr-182. CTGF is unstable in vivo, being converted into low molecular mass forms (10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) by an unidentified protease (42). Interestingly, conditioned medium from serum-stimulated mouse fibroblasts did not degrade CTGF (43), suggesting that a cell surface protease was responsible for CTGF processing.…”

“…Nonetheless, because proteases do not operate alone but more commonly in amplification cascades or regulatory circuits in the presence of a multitude of interacting proteins, substrates, and cleavage products, and often in distinct compartments, biologically relevant protease substrates in vivo may differ from theoretical activities inferred from in vitro experiments. In the face of such biochemical complexity, this emphasizes the need to identify protease-cleaved substrates and not just enzyme activities in cells, tissues, and whole organisms (10,11). Thus, rapid techniques are needed to directly identify new substrates in biological settings and to quantitate differences in substrate processing as disease biomarkers and as surrogate markers of antiproteolytic drug treatment (3).…”

Design and Synthesis of Sulfur Based Inhibitors of Matrix Metalloproteinase-1.

“…In these physiological processes, MMP activity is tightly regulated. [1][2][3][4][5] However, excessive MMP synthesis and release can lead to connective tissue degradation and destruction, which occurs in tumor invasion, metastasis, 6) corneal ulceration, 7) arthritis, 8a) periodontal disease, 9) and multiple sclerosis. 10) A number of MMP inhibitors have progressed into clinical trials mainly for cancer and rheumatoid arthritis.…”

Novel Hydroquinone as a Matrix Metallo-proteinase Inhibitor from the Mushroom,Piptoporus betulinus

“…In addition, MMPs have been implicated in several pathological processes including arthritis, tumor growth, metastasis, periodontal disease, and multiple sclerosis. [2][3][4][5][6] Therefore, the discovery of potent inhibitors of these enzymes is a highly attractive endeavor both scientiˆcally and commercially.…”