Biochemical Characterization of the Catalytic Domain of Human Matrix Metalloproteinase 19

Abstract

We have recently cloned MMP-19, a novel matrix metalloproteinase, which, due to unique structural features, was proposed to represent the first member of a new MMP subfamily (Pendá s, A. M., Knä uper, V., Puente, X. S., Llano, E., Mattei, M. G., Apte, S., Murphy, G., and Ló pez-Otin, C. -Pro-Leu-Gly-Dpa-Ala-Arg-NH 2 and, with higher efficiency, the stromelysin substrate McaPro-Leu-Ala-Nva-Dpa-Ala-Arg-NH 2 . Kinetic analysis of the interactions of the catalytic domain of MMP-19 with the natural MMP inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), showed strong inhibition using TIMP-2, TIMP-3, and TIMP-4, while TIMP-1 was less efficient. We also demonstrated that synthetic hydroxamic acid-based compounds efficiently inhibited the enzyme. The catalytic domain of MMP-19 was able to hydrolyze the basement membrane components type IV collagen, laminin, and nidogen, as well as the large tenascin-C isoform, fibronectin, and type I gelatin in vitro, suggesting that MMP-19 is a potent proteinase capable of hydrolyzing a broad range of extracellular matrix components. Neither the catalytic domain nor the fulllength MMP-19 was able to degrade triple-helical collagen. Finally, and in contrast to studies with other MMPs, MMP-19 catalytic domain was not able to activate any of the latent MMPs tested in vitro.

Method Development of Efficient Protein Extraction in Bone Tissue for Proteome Analysis

“…38 MMP-2, also previously known as the Type IV collagenase, 39 had substrate specificity to denature collagens, Type V, VII, X, XII collagens, vitronectin, aggrecan, galectin-3, and elastin, and most of them were also identified. Various substrates of MMP-19 have been identified in vitro 40 , including type IV collagen, gelatin, laminin-1, nidogen 1, fibronectin, and tenascin-C, which were also mostly detected. The identification of these bone-specific proteins demonstrated the effective of the sequential protein extraction protocol developed in this study.…”

Matrix Metalloproteinase Inhibition Decreases Ischemia-Reperfusion Injury After Lung Transplantation

“…Accordingly, an in vivo study by Betsuyaku et al have suggested that neutrophil emigration in LPS-injured lungs occurs in MMP9-deficient mice, and that this molecule is thus not necessary for the neutrophils to cross the basement membrane (56). Pathophysiological differences existing between lung IR injury and LPS-induced lung injury, non-identified compensatory mechanisms of MMP9-deficient mice, and the involvement of other MMPs, like MMP2, membrane-type-MMP or the recently characterized MMP19, thought to play a significant role for basement membrane disruption (57), may account for the apparent discrepancy between the various experimental studies.…”

Matrix metalloproteinase‐19 is expressed by proliferating epithelium but disappears with neoplastic dedifferentiation

“…3,8 In vitro, MMP-19 is able to degrade many important BM components, e.g., type IV collagen, laminin-1, nidogen, fibronectin, tenascin-C isoform, aggrecan, type I gelatin and cartilage oligomeric matrix protein, 9 but does not activate any other latent MMPs. 10 By Northern hybridization, MMP-19 was originally detected in placenta, lung, pancreas, liver, ovary, spleen and intestine. 7 Independently, it was isolated as an autoantigen from the inflamed synovium of a patient suffering from rheumatoid arthritis.…”

“…11,13 Because of its appearance in normal tissues, it is possible that MMP-19 is important in normal tissue remodeling or activation of secreted and membrane-bound proteins, like growth factors. 10 Our aim was to determine whether MMP-19 is induced in the epithelium during remodeling associated with either wound repair or cancer invasion. We show here that MMP-19 can be expressed by proliferating, but not migrating, keratinocytes; is upregulated by TNF-␣ and PMA in keratinocyte cultures; and, unlike several classical MMPs, disappears from proliferating keratinocytes during neoplastic dedifferentiation.…”