title: In vitro reconstitution of complexes between pro-matrix metalloproteinase-9 and the proteoglycans serglycin and versican.
abstract: Previously, we have shown that a proportion of the matrix metalloproteinase-9 (MMP-9) synthesized by the macrophage cell line THP-1 binds to a chondroitin sulfate proteoglycan (CSPG) core protein to form a reduction-sensitive heteromer. It was also shown that the hemopexin-like (PEX) domain and the fibronectin-like (FnII) module in the enzyme are involved in heteromer formation. In this paper, we show that reduction-sensitive and SDS-stable heteromers may be reconstituted in vitro by mixing proMMP-9 with either serglycin, versican or CSPGs isolated from various monocytic cell lines. In addition, a strong but SDS-soluble proMMP-9·CSPG heteromer was formed. The two macromolecules in the SDS-stable reduction-sensitive heteromers were not linked together by disulfide bonds. As for the heteromer isolated from THP-1 cells, in vitro reconstituted SDS-stable and SDS-soluble heteromers showed weaker binding to gelatin than the proMMP-9 monomer. Furthermore, gelatin inhibited in vitro reconstitution of the heteromers, showing that the FnII module is involved in the complex formation. Tissue inhibitor of metalloproteinase (TIMP)-1 was not be detected in the proMMP-9·CSPG complexes. However, the presence of TIMP-1 inhibited formation of the SDS-soluble heteromer, but not the SDS-stable reduction-sensitive heteromer. This indicates that different regions in the PEX domain are involved formation of these heteromers.
authors.affiliation: Department of Medical Biology, University of Tromsø, Norway.
authors.forename: Nabin
authors.initials: N
authors.lastname: Malla
authors.forename: Eli
authors.initials: E
authors.lastname: Berg
authors.forename: Achilleas D
authors.initials: AD
authors.lastname: Theocharis
authors.forename: Gunbjørg
authors.initials: G
authors.lastname: Svineng
authors.forename: Lars
authors.initials: L
authors.lastname: Uhlin-Hansen
authors.forename: Jan-Olof
authors.initials: JO
authors.lastname: Winberg
mesh_terms: D045744:Cell Line, Tumor; D002809:Chondroitin Sulfates / Q000737:chemistry*; D003553:Cystine / Q000737:chemistry; D003902:Detergents / Q000737:chemistry; D004792:Enzyme Precursors / Q000737:chemistry*; D004795:Enzyme Stability; D005780:Gelatin / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D020780:Matrix Metalloproteinase 9 / Q000737:chemistry*; D046912:Multiprotein Complexes / Q000737:chemistry; D017830:Octoxynol / Q000737:chemistry; D011485:Protein Binding; D011509:Proteoglycans / Q000737:chemistry*; D012965:Sodium Chloride / Q000737:chemistry; D012967:Sodium Dodecyl Sulfate / Q000737:chemistry; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry; D053675:Versicans / Q000737:chemistry*; D033921:Vesicular Transport Proteins / Q000737:chemistry*
chemical_list: D003902:Detergents; D004792:Enzyme Precursors; D046912:Multiprotein Complexes; D011509:Proteoglycans; C560822:TIMP1 protein, human; D019715:Tissue Inhibitor of Metalloproteinase-1; C508322:VCAN protein, human; D033921:Vesicular Transport Proteins; C075430:serglycin; D053675:Versicans; D012967:Sodium Dodecyl Sulfate; D012965:Sodium Chloride; D003553:Cystine; D005780:Gelatin; D017830:Octoxynol; D002809:Chondroitin Sulfates; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9