Matrilysin Is Much More Efficient Than Other Matrix Metalloproteinases in the Proteolytic Inactivation of α1-Antitrypsin

Macrophage elastase (ME) was originally named when metal-dependent elastolytic activity was detected in conditioned media of murine macrophages. Subsequent cDNA cloning of the mouse and human enzyme demonstrated that ME is a distinct member of the matrix metalloproteinase family. To date, the catalytic parameters that describe the hydrolysis of elastin by ME have not been quantified and its activity against other matrix proteins have not been described. In this report, we have examined the action of purified recombinant human ME (rHME), produced in Escherichia coli, on elastin and other extracellular matrix proteins. On a molar basis, rHME is approximately 30% as active as human leukocyte elastase in solubilizing elastin. rHME also efficiently degrades ␣ 1 -antitrypsin (␣ 1 -AT), the primary physiological inhibitor of human leukocyte elastase. In addition, rHME efficiently degrades fibronectin, laminin, entactin, type IV collagen, chondroitan sulfate, and heparan sulfate. These results suggest that HME may be required for macrophages to penetrate basement membranes and remodel injured tissue during inflammation. Moreover, abnormal expression of HME may contribute to destructive processes such as pulmonary emphysema and vascular aneurysm formation. To further understand the specificity of HME, the initial cleavage sites in ␣ 1 -AT have been determined. In addition, the hydrolysis of a series of synthetic peptides with different P 1 residues has been determined. rHME can accept large and small amino acids at the P 1 site, but has a preference for leucine.Macrophage elastase (MMP-12) shares many properties with other members of the matrix metalloproteinase (MMP) 1 gene family, yet it is unique in several ways. Like other MMPs, metalloelastase requires zinc for catalytic activity, is inhibited by the tissue inhibitors of metalloproteinases (TIMPs), and has common structural domains with other MMPs (1, 2). Human macrophage elastase (HME) is most closely related to collagenase-1 (MMP-1) and stromelysin-1 (MMP-3), being 49% identical to each at the amino acid level (3). Moreover, the gene for macrophage elastase, composed of a common 10-exon, 9-intron structure, is on human chromosome 11q22.2/22.3 with at least six other MMPs (4). Despite these similarities, HME possesses certain distinct biological and biochemical properties. Expression appears to be largely restricted to tissue macrophages (4). Upon activation, it not only cleaves its 8-kDa N-terminal domain, but also has a unique propensity to autolytically release its 23-kDa C-terminal domain resulting in a mature active 22-kDa proteinase (3)(4)(5).Macrophage elastase shares its elastolytic activity (6, 7) with only a few MMPs, including the gelatinases (MMP-2 and MMP-9) and, to a lesser extent, matrilysin (8, 9). However, despite this characteristic activity, the relative capacity of metalloelastase (human or mouse) to degrade elastin has never been quantified. In addition, the catalytic capacity of metalloelastase against other extracellular matrix components has...

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“…Matrilysin expressed in bacteria had equal catalytic activity to matrilysin expressed in eukaryotic cells (12).…”

Section: Methodsmentioning

confidence: 94%

“…The primary physiological inhibitor is ␣ 1 -AT which forms a stable complex with the enzyme (19). A variety of MMPs are capable of degrading ␣ 1 -AT (12,15,16) and abolishing its ability to inhibit HLE. MMPs, therefore, may significantly modulate the physiological role of ␣ 1 -AT.…”

Section: Hme and Elastolysis 12192mentioning

confidence: 99%

Hydrolysis of a Broad Spectrum of Extracellular Matrix Proteins by Human Macrophage Elastase

Gronski

Martin²,

Kobayashi

et al. 1997

Journal of Biological Chemistry

275

204

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“…In vitro studies have demonstrated that reactive oxygen species released by activated leukocytes can inactivate serpins (␣ 1 -proteinase inhibitor and SLPI) as well as ␣ 2 -macroglobulin [98][99][100][101]. Serpins are also susceptible to proteolytic inactivation by several classes of proteinases: (1) several MMPs (MMP-1, MMP-3, MMP-7, MMP-8, MMP-9, and MMP-12) can inactivate ␣ 1 -proteinase inhibitor, ␣ 1 -antichymotrypsin, and also ␣ 2 -macroglobulin [25,26,[102][103][104]; (2) the serine proteinase, HLE, can inactivate its cognate inhibitor (␣ 1 -proteinase inhibitor) [105] and PAI-1 [106]; (3) cathepsin B (a cysteine proteinase) can inactivate ␣ 1 -proteinase inhibitor [107]; and (4) several bacterial proteinases can inactivate ␣ 1 -proteinase inhibitor and SLPI [108]. It is also noteworthy that serine proteinases can proteolytically inactivate TIMPs [109].…”

Section: Inactivation Of Proteinase Inhibitorsmentioning

confidence: 99%

The cell biology of leukocyte-mediated proteolysis

Owen

Campbell

1999

Journal of Leukocyte Biology

383

399

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Leukocyte-derived proteinases have the capacity to degrade every component of the extracellular matrix, and thereby play fundamental roles in physiological processes. However, if the activity of these proteinases is uncontrolled or dysregulated, they have the capacity to contribute to tissue injury that potentially affects every organ in the body. Although there is a substantial literature on structure and activity of these proteinases when they are free in solution, until recently there has been little information about the cell biology of proteinases and their inhibitors. Recent studies, however, have identified several mechanisms by which inflammatory cells can degrade extracellular proteins in a milieu that contains high-affinity proteinase inhibitors. J. Leukoc. Biol. 65: 137-150; 1999.

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“…Activation of the proenzyme involves proteolytic removal of the N-terminal proregion containing the cysteine switch motif conserved in matrix metalloproteinases (18 ). MMP-7 can cleave a broad range of extracellular matrix macromolecules such as fibronectin, laminin, proteoglycan, elastin, gelatin, and type IV collagen as well as ␣ 1 -antitrypsin (19,20 ). Overexpression of MMP-7 has been reported in several cancerous tissues, such as colorectal, ovarian, and pancreatic cancer (21)(22)(23).…”

mentioning

confidence: 99%

Identification of Pro-MMP-7 as a Serum Marker for Renal Cell Carcinoma by Use of Proteomic Analysis

Sarkissian¹,

Fergelot

Lamy

et al. 2008

Clinical Chemistry

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Background: No validated renal cell carcinoma (RCC) marker is known for detection of asymptomatic disease in selected populations or for prognostic purposes or treatment monitoring. We identified immunogenic proteins as tumor markers for RCC by combining conventional proteome analysis with serological screening, and we investigated the diagnostic clinical value of such markers in serum. Methods: We studied the immunogenic protein expression profile of CAL 54, a human RCC cell line, by 2-dimensional electrophoresis combined with immunoblotting using sera from healthy donors compared with RCC patients. We developed a homogeneous, fluorescent, dual-monoclonal immunoassay for metalloproteinase 7 (MMP-7) and used it to measure MMP-7 in sera from 30 healthy donors, 30 RCC patients, and 40 control patients. Results: Pro-MMP-7 (29 kDa; pI 7.7) in the CAL 54 cell line secretome was an immunogenic protein reactive with RCC patient sera but not with control sera. The concentrations of pro-MMP-7 were increased (P <0.0001) in sera of RCC patients (median 7.56 μg/L; range 3.12–30.5 μg/L) compared with healthy controls (median 2.13 μg/L; range 0.17–3.5 μg/L). Serum pro-MMP-7 had a sensitivity of 93% (95% CI 78%–99%) at a specificity of 75% (59%–87%) for RCC in the samples tested. Conclusion: Proteomics technology combined with serology led to the identification of serum pro-MMP-7 as a marker of RCC and represents a powerful tool in searching for candidate proteins as biomarkers.

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Matrilysin Is Much More Efficient Than Other Matrix Metalloproteinases in the Proteolytic Inactivation of α1-Antitrypsin

Cited by 114 publications

References 0 publications

Hydrolysis of a Broad Spectrum of Extracellular Matrix Proteins by Human Macrophage Elastase

Hydrolysis of a Broad Spectrum of Extracellular Matrix Proteins by Human Macrophage Elastase

The cell biology of leukocyte-mediated proteolysis

Identification of Pro-MMP-7 as a Serum Marker for Renal Cell Carcinoma by Use of Proteomic Analysis

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