Clusterin, an abundant serum factor, is a possible negative regulator of MT6-MMP/MMP-25 produced by neutrophils

J Biol Chem. 2003 Sep 19;278(38):36350-7. doi: 10.1074/jbc.M301509200. Epub 2003 Jul 14.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

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

Substances

  • CLU protein, human
  • Clusterin
  • DNA, Complementary
  • Epitopes
  • GPI-Linked Proteins
  • Glycoproteins
  • Molecular Chaperones
  • Recombinant Proteins
  • Furin
  • Matrix Metalloproteinases
  • Matrix Metalloproteinases, Membrane-Associated
  • matrix metalloproteinase 25