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 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.
Mysliwy
Justyna
J
Biochemisches Institut der Christian-Albrechts-Universität Kiel, Olshausenstr. 40 24118 Kiel, Germany.
Dingley
Andrew J
AJ
Sedlacek
Radislav
R
Grötzinger
Joachim
J
D049109:Cell Proliferation / Q000187:drug effects; D006801:Humans; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000737:chemistry; D007334:Insulin-Like Growth Factor I / Q000737:chemistry; D015603:Keratinocytes / Q000166:cytology / Q000502:physiology; D053505:Matrix Metalloproteinases, Secreted; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology; D011485:Protein Binding / Q000235:genetics; D017434:Protein Structure, Tertiary / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship; D020349:Surface Plasmon Resonance
D018972:Insulin-Like Growth Factor Binding Protein 3; D011994:Recombinant Proteins; D007334:Insulin-Like Growth Factor I; D053505:Matrix Metalloproteinases, Secreted; D008666:Metalloendopeptidases; C108362:matrix metalloproteinase 19