Syndecan-1 interaction with the LG4/5 domain in laminin-332 is essential for keratinocyte migration

J Cell Physiol. 2008 Jan;214(1):238-49. doi: 10.1002/jcp.21184.

Abstract

Laminin 5/laminin 332 (LN332) is an adhesion substrate for epithelial cells. After secretion of LN332, a regulated cleavage occurs at the carboxy-terminus of its alpha3 subunit, which releases a tandem of two globular modules named LG4/5. We show that the presence of the LG4/5 domain in precursor LN332 decreases its integrin-mediated cell adhesion properties in comparison with mature LN332. Whereas cell adhesion to the recombinant LG4/5 fragment relies solely on the heparan sulfate proteoglycan (HSPG) receptor syndecan-1, we reveal that both syndecan-1 and the alpha3beta1 integrin bind to precursor LN332. We further demonstrate that syndecan-1 mediated cell adhesion to the LG4/5 fragment and pre-LN332 allows the formation of fascin-containing protrusions, depending on the GTPases Rac and Cdc42 activation. Reducing syndecan-1 expression in normal keratinocytes prevents cell protrusions on pre-LN332 with subsequent failure of the peripheral localization of the alpha3beta1 integrin. We finally show that cell migration on pre-LN332 requires syndecan-1. Therefore, the LG4/5 domain in precursor LN332 appears to trigger intracellular signaling events, which participate in keratinocyte motility.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Adhesion
  • Cell Adhesion Molecules / chemistry*
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / isolation & purification
  • Cell Adhesion Molecules / metabolism*
  • Cell Movement / physiology*
  • Cells, Cultured
  • Enzyme Activation
  • Fibrosarcoma / pathology
  • Fluorescein-5-isothiocyanate
  • Fluorescent Dyes
  • Humans
  • Integrin alpha3beta1 / metabolism
  • Kalinin
  • Keratinocytes / physiology*
  • Male
  • Melanoma / pathology
  • Microscopy, Video
  • Phalloidine
  • Plasmids
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Rhodamines
  • Skin / cytology
  • Syndecan-1 / metabolism*
  • Transfection
  • cdc42 GTP-Binding Protein / analysis
  • cdc42 GTP-Binding Protein / metabolism
  • rac GTP-Binding Proteins / analysis
  • rac GTP-Binding Proteins / metabolism

Substances

  • Cell Adhesion Molecules
  • Fluorescent Dyes
  • Integrin alpha3beta1
  • Recombinant Proteins
  • Rhodamines
  • SDC1 protein, human
  • Syndecan-1
  • Phalloidine
  • cdc42 GTP-Binding Protein
  • rac GTP-Binding Proteins
  • Fluorescein-5-isothiocyanate