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MT1-MMP shedding involves an ADAM and is independent of its localization in lipid rafts

Marta Toth1, Anjum Sohail, Shahriar Mobashery

  • 1Department of Chemistry and Biochemistry and the Walther Cancer Research Center, University of Notre Dame, Notre Dame, IN 46556, USA.

Biochemical and Biophysical Research Communications
|September 30, 2006
PubMed

Insights

The shedding of membrane type 1-matrix metalloproteinase (MT1-MMP) occurs in two steps via integral membrane proteases. Differential inhibition suggests ADAM proteases mediate the final cleavage, releasing soluble MT1-MMP forms.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Proteomics

Background:

  • Membrane type 1-matrix metalloproteinase (MT1-MMP) is a cell surface protease involved in pericellular proteolysis.
  • The ectodomain of MT1-MMP can be shed, but the mechanisms and resulting soluble forms are not fully understood.

Purpose of the Study:

  • To elucidate the mechanism and identify the proteases involved in MT1-MMP ectodomain shedding.
  • To characterize the soluble forms of MT1-MMP released from HT1080 cells.

Main Methods:

  • Analysis of soluble MT1-MMP forms in HT1080 cells using biochemical assays.
  • Investigation of the shedding process in purified plasma membranes.
  • Assessment of the role of lipid rafts and inhibition studies using TIMP-3 and potential ADAM inhibitors.

Main Results:

  • MT1-MMP is shed as two soluble forms (~52 kDa and ~50 kDa) through a two-step, time-dependent process.
  • Integral membrane metalloproteases mediate the release of these soluble species.
  • The second cleavage step, forming the 50 kDa species, is likely mediated by an ADAM protease, indicated by differential TIMP-3 sensitivity.
  • Shedding is independent of MT1-MMP's lipid raft localization.

Conclusions:

  • MT1-MMP ectodomain shedding is a regulated, multi-step proteolytic process.
  • ADAM proteases play a role in generating specific soluble MT1-MMP forms.
  • Understanding MT1-MMP shedding provides insights into regulating pericellular proteolysis.

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