Identification of CD63 as a tissue inhibitor of metalloproteinase-1 interacting cell surface protein

EMBO J. 2006 Sep 6;25(17):3934-42. doi: 10.1038/sj.emboj.7601281. Epub 2006 Aug 17.


This study identified CD63, a member of the tetraspanin family, as a TIMP-1 interacting protein by yeast two-hybrid screening. Immunoprecipitation and confocal microscopic analysis confirmed CD63 interactions with TIMP-1, integrin beta1, and their co-localizations on the cell surface of human breast epithelial MCF10A cells. TIMP-1 expression correlated with the level of active integrin beta1 on the cell surface independent of cell adhesion. While MCF10A cells within a three-dimensional (3D) matrigel matrix form polarized acinar-like structures, TIMP-1 overexpression disrupted breast epithelial cell polarization and inhibited caspase-mediated apoptosis in centrally located cells, necessary for the formation and maintenance of the hollow acinar-like structures. Small hairpin RNA (shRNA)-mediated CD63 downregulation effectively reduced TIMP-1 binding to the cell surface, TIMP-1 co-localization with integrin beta1, and consequently reversed TIMP-1-mediated integrin beta1 activation, cell survival signaling and apoptosis inhibition. CD63 downregulation also restored polarization and apoptosis of TIMP-1 overexpressing MCF10A cells within a 3D-matrigel matrix. Taken together, the present study identified CD63 as a cell surface binding partner for TIMP-1, regulating cell survival and polarization via TIMP-1 modulation of tetraspanin/integrin signaling complex.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antigens, CD / metabolism
  • Antigens, CD / physiology*
  • Apoptosis
  • Cell Adhesion
  • Cell Line
  • Cell Membrane / metabolism
  • Cell Polarity
  • Cell Survival
  • Down-Regulation
  • Epithelial Cells / cytology
  • Epithelial Cells / metabolism
  • Gene Library
  • Humans
  • Integrin beta1 / metabolism*
  • Mammary Glands, Human / cytology
  • Membrane Proteins / metabolism*
  • Platelet Membrane Glycoproteins / metabolism
  • Platelet Membrane Glycoproteins / physiology*
  • Protein Binding
  • Signal Transduction
  • Tetraspanin 30
  • Tissue Inhibitor of Metalloproteinase-1 / metabolism*
  • Two-Hybrid System Techniques


  • Antigens, CD
  • CD63 protein, human
  • Integrin beta1
  • Membrane Proteins
  • Platelet Membrane Glycoproteins
  • Tetraspanin 30
  • Tissue Inhibitor of Metalloproteinase-1