Glycosylation broadens the substrate profile of membrane type 1 matrix metalloproteinase

J Biol Chem. 2004 Feb 27;279(9):8278-89. doi: 10.1074/jbc.M311870200. Epub 2003 Dec 11.


Membrane type 1 matrix metalloproteinase (MT1-MMP) is a collagenolytic enzyme that has been implicated in normal development and in pathological processes such as cancer metastasis. The activity of MT1-MMP is regulated extensively at the post-translational level, and the current data support the hypothesis that MT1-MMP activity is modulated by glycosylation. Enzymatic deglycosylation, site-directed mutagenesis, and lectin precipitation assays were used to demonstrate that MT1-MMP contains O-linked complex carbohydrates on the Thr(291), Thr(299), Thr(300), and/or Ser(301) residues in the proline-rich linker region. MT1-MMP glycoforms were detected in human cancer cell lines, suggesting that MT1-MMP activity may be regulated by differential glycosylation in vivo. Although the autolytic processing and interstitial collagenase activity of MT1-MMP were not impaired in glycosylation-deficient mutants, cell surface MT1-MMP-catalyzed activation of pro-matrix metalloproteinase-2 (proMMP-2) required proper glycosylation of MT1-MMP. The inability of carbohydrate-free MT1-MMP to activate proMMP-2 was not a result of defective MT1-MMP zymogen activation, aberrant protein stability, or inability of the mature enzyme to oligomerize. Rather, our data support a mechanism whereby glycosylation affects the recruitment of tissue inhibitor of metalloproteinases-2 (TIMP-2) to the cell surface, resulting in defective formation of the MT1-MMP/TIMP-2/proMMP-2 trimeric activation complex. These data provide evidence for an additional mechanism for post-translational control of MT1-MMP activity and suggest that glycosylation of MT1-MMP may regulate its substrate targeting.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Carbohydrate Conformation
  • Cell Membrane / metabolism
  • Chemical Precipitation
  • Chlorocebus aethiops
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Activation
  • Glycoside Hydrolases / metabolism
  • Glycosylation
  • Humans
  • Lectins
  • Matrix Metalloproteinase 2 / metabolism
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases / chemistry*
  • Metalloendopeptidases / genetics
  • Metalloendopeptidases / metabolism*
  • Molecular Sequence Data
  • Mutagenesis
  • Mutagenesis, Site-Directed
  • Protein Folding
  • Protein Precursors / metabolism
  • Protein Processing, Post-Translational
  • Structure-Activity Relationship
  • Substrate Specificity
  • Tissue Inhibitor of Metalloproteinase-2 / metabolism
  • Tumor Cells, Cultured


  • Lectins
  • Protein Precursors
  • Tissue Inhibitor of Metalloproteinase-2
  • Glycoside Hydrolases
  • Matrix Metalloproteinases, Membrane-Associated
  • Metalloendopeptidases
  • Matrix Metalloproteinase 2