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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.

Type

Journal article

Journal

The Journal of biological chemistry

Publication Date

09/2003

Volume

278

Pages

36350 - 36357

Addresses

Division of Cancer Cell Research, Institute of Medical Science, the University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.

Keywords

Neutrophils, Cell Line, COS Cells, Cell Line, Tumor, Animals, Dogs, Humans, Matrix Metalloproteinases, Furin, Glycoproteins, Molecular Chaperones, Recombinant Proteins, DNA, Complementary, Epitopes, Blotting, Western, Precipitin Tests, Chromatography, Gel, Electrophoresis, Polyacrylamide Gel, Silver Staining, Transfection, Gene Expression Regulation, Amino Acid Motifs, Catalytic Domain, Protein Structure, Tertiary, Protein Binding, Dose-Response Relationship, Drug, Kinetics, Genetic Vectors, Plasmids, Hydrogen-Ion Concentration, Models, Genetic, Time Factors, Clusterin, Matrix Metalloproteinases, Membrane-Associated, GPI-Linked Proteins