Structural characterization and binding properties of the hemopexin-like domain of the matrixmetalloproteinase-19

The matrixmetalloproteinase-19 (MMP-19) belongs to the superfamily of the zinc-dependent endopeptidases, which are secreted by cells and are involved in the remodeling of the extracellular matrix. The full-length protein consists of a signal peptide, a propeptide, a catalytic domain and a C-terminal...

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Bibliographic Details
Published in:Protein expression and purification 2006-04, Vol.46 (2), p.406-413
Main Authors: Mysliwy, Justyna, Dingley, Andrew J., Sedlacek, Radislav, Grötzinger, Joachim
Format: Article
Language:English
Subjects:
Cell Proliferation - drug effects
Expression
Hemopexin domain
Humans
IGFBP-3
Insulin-Like Growth Factor Binding Protein 3 - chemistry
Insulin-Like Growth Factor I - chemistry
Keratinocytes - cytology
Keratinocytes - physiology
Matrix Metalloproteinases, Secreted
Matrixmetalloproteinase-19
Metalloendopeptidases - chemistry
Metalloendopeptidases - genetics
Metalloendopeptidases - pharmacology
NMR
Protein Binding - genetics
Protein Structure, Tertiary - genetics
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Refolding
Structure-Activity Relationship
Surface Plasmon Resonance
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Summary:The matrixmetalloproteinase-19 (MMP-19) belongs to the superfamily of the zinc-dependent endopeptidases, which are secreted by cells and are involved in the remodeling of the extracellular matrix. The full-length protein consists of a signal peptide, a propeptide, a catalytic domain and a C-terminal hemopexin-like domain. For other members of this superfamily, the hemopexin-like domain has been described to be involved in substrate recognition. In this study, the hemoxpexin domain of MMP-19 was expressed in Escherichia coli, refolded, and purified. For structural characterization, circular dichroism and NMR spectroscopy were used. We show that the hemopexin-like domain of MMP-19 is able to bind calcium and this binding induces a conformational change and an increase in the thermal stability of the domain. MMP-19 promotes proliferation of keratinocytes by cleaving the insulin-like-growth factor binding protein-3, thereby causing the release of IGF-1, which is a potent growth factor for these cells. By plasmon resonance experiments, we show that the isolated hemopexin-like domain is able to bind to the insulin-like-growth factor binding protein-3. These results provide a basis for further structural investigations that could be used for the rational design of potential agonists and antagonists.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2005.08.020