Clusterin, an abundant serum factor, is a possible negative regulator of MT6-MMP/MMP-25 produced by neutrophils.
MT6-MMP/MMP-25 is the latest member of the membrane-type matrix metalloproteinase (MT-MMP) subgroup in the MMP family and is expressed in neutrophils and some brain tumors. The proteolytic activity of MT6-MMP has been studied using recombinant catalytic fragments and shown to degrade several components of the extracellular matrix. However, the activity ... is possibly modulated further by the C-terminal hemopexin-like domain, because some MMPs are known to interact with other proteins through this domain. To explore the possible function of this domain, we purified a recombinant MT6-MMP with the hemopexin-like domain as a soluble form using a Madin-Darby canine kidney cell line as a producer. Mature and soluble MT6-MMP processed at the furin motif was purified as a 45-kDa protein together with a 46-kDa protein having a single cleavage in the hemopexin-like domain. Interestingly, 73- and 70-kDa proteins were co-purified with the soluble MT6-MMP by forming stable complexes. They were identified as clusterin, a major component of serum, by N-terminal amino acid sequencing. MT1-MMP that also has a hemopexin-like domain did not form a complex with clusterin. MT6-MMP forming a complex with clusterin was detected in human neutrophils as well. The enzyme activity of the soluble MT6-MMP was inactive in the clusterin complex. Purified clusterin was inhibitory against the activity of soluble MT6-MMP. On the other hand, it had no effect on the activities of MMP-2 and soluble MT1-MMP. Because clusterin is an abundant protein in the body fluid in tissues, it may act as a negative regulator of MT6-MMP in vivo.
Mesh Terms:
Amino Acid Motifs, Animals, Blotting, Western, COS Cells, Catalytic Domain, Cell Line, Cell Line, Tumor, Chromatography, Gel, Clusterin, DNA, Complementary, Dogs, Dose-Response Relationship, Drug, Electrophoresis, Polyacrylamide Gel, Epitopes, Furin, Gene Expression Regulation, Genetic Vectors, Glycoproteins, Humans, Hydrogen-Ion Concentration, Kinetics, Matrix Metalloproteinases, Matrix Metalloproteinases, Membrane-Associated, Models, Genetic, Molecular Chaperones, Neutrophils, Plasmids, Precipitin Tests, Protein Binding, Protein Structure, Tertiary, Recombinant Proteins, Silver Staining, Time Factors, Transfection
Amino Acid Motifs, Animals, Blotting, Western, COS Cells, Catalytic Domain, Cell Line, Cell Line, Tumor, Chromatography, Gel, Clusterin, DNA, Complementary, Dogs, Dose-Response Relationship, Drug, Electrophoresis, Polyacrylamide Gel, Epitopes, Furin, Gene Expression Regulation, Genetic Vectors, Glycoproteins, Humans, Hydrogen-Ion Concentration, Kinetics, Matrix Metalloproteinases, Matrix Metalloproteinases, Membrane-Associated, Models, Genetic, Molecular Chaperones, Neutrophils, Plasmids, Precipitin Tests, Protein Binding, Protein Structure, Tertiary, Recombinant Proteins, Silver Staining, Time Factors, Transfection
J. Biol. Chem.
Date: Sep. 19, 2003
PubMed ID: 12860995
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