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Macrophage metalloelastase degrades matrix and myelin proteins and processes a tumour necrosis factor-alpha fusion
S Chandler1, J Cossins, J Lury
1British Biotech Pharmaceuticals Ltd., Cowley, Oxford, UK. chandler@britbio.co.uk
Abstract:
The matrix metalloproteinases (MMPs) are a group of enzymes which have the ability to degrade extracellular matrix. They also cleave non-matrix proteins such as myelin basic protein and alpha 1-antitrypsin and they are able to process tumour necrosis factor-alpha (TNF) to its mature form. We have cloned, expressed and purified human macrophage metalloelastase (EC 3.4.24.65), an MMP recognised for its ability to degrade elastin, but whose substrate specificity has not yet been defined. With the exception of type I collagen this enzyme degraded all matrix proteins tested, namely: type IV collagen, type I gelatin, fibronectin, laminin, vitronectin and proteoglycan. It also degraded myelin basic protein, cleaved alpha 1-antitrypsin and released TNF from a pro-TNF fusion protein. Thus, in common with several other MMPs, macrophage metalloelastase has a broad substrate range which extends beyond that of elastin alone.
Insights
Matrix metalloproteinases (MMPs) are enzymes that degrade extracellular matrix. Human macrophage metalloelastase, an MMP, was studied and found to degrade a broad range of matrix and non-matrix proteins beyond just elastin.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Biology
Background:
- Matrix metalloproteinases (MMPs) are enzymes crucial for extracellular matrix (ECM) remodeling.
- MMPs also process non-matrix substrates, including cytokines like tumor necrosis factor-alpha (TNF).
- Human macrophage metalloelastase (hMME) is an MMP known for elastin degradation, but its substrate specificity is not fully defined.
Purpose of the Study:
- To clone, express, and purify human macrophage metalloelastase (hMME).
- To determine the substrate specificity of hMME beyond elastin degradation.
- To investigate the enzymatic activity of hMME on various matrix and non-matrix proteins.
Main Methods:
- Cloning and expression of human macrophage metalloelastase.
- Purification of the recombinant enzyme.
- Enzymatic assays using a panel of matrix proteins (type IV collagen, gelatin, fibronectin, laminin, vitronectin, proteoglycan) and non-matrix proteins (myelin basic protein, alpha 1-antitrypsin, pro-TNF fusion protein).
Main Results:
- Human macrophage metalloelastase was successfully cloned, expressed, and purified.
- The enzyme degraded all tested matrix proteins except type I collagen.
- hMME also degraded myelin basic protein, cleaved alpha 1-antitrypsin, and processed pro-TNF to its mature form.
Conclusions:
- Human macrophage metalloelastase possesses a broad substrate range.
- Its enzymatic activity extends beyond elastin to include various ECM components and non-matrix proteins.
- These findings highlight the diverse roles of hMME in biological processes.