The actin cytoskeleton regulates LFA-1 ligand binding through avidity rather than affinity changes

Y van Kooyk, S J van Vliet, C G Figdor

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132 Citations (Scopus)


To elucidate the role of the cytoskeleton regulating avidity or affinity changes in the leukocyte adhesion receptor lymphocyte function-associated antigen-1 (LFA-1) (alpha(L)beta(2)), we generated mutant cytoplasmic LFA-1 receptors and expressed these into the erythroleukemic cell line K562. We determined whether intercellular adhesion molecule-1 (ICAM-1)-mediated adhesion of LFA-1, lacking parts of its cytoplasmic tails, is regulated through receptor diffusion/clustering and/or by altered ligand binding affinity. All cytoplasmic deletion mutants that lack the complete beta(2) cytoplasmic tail and/or the conserved KVGFFKR sequence in the alpha(L) cytoplasmic tail were constitutively active and expressed high levels of the activation epitopes NKI-L16 and M24. Surprisingly, whereas these mutants showed a clustered cell surface distribution of LFA-1, the ligand-binding affinity as measured by titration of soluble ligand ICAM-1 remained unaltered. The notion that redistribution of LFA-1 does not alter ligand-binding affinity is further supported by the finding that disruption of the cytoskeleton by cytochalasin D did not alter the binding affinity nor adhesion to ICAM-1 of these mutants. Most cytoplasmic deletion mutants that spontaneously bound ICAM-1 were not capable to spread on ICAM-1, demonstrating that on these mutants LFA-1 is not coupled to the actin cytoskeleton. From these data we conclude that LFA-1-mediated cell adhesion to ICAM-1 is predominantly regulated by receptor clustering and that affinity alterations do not necessarily coincide with strong ICAM-1 binding.

Original languageEnglish
Pages (from-to)26869-77
Number of pages9
JournalJournal of Biological Chemistry
Issue number38
Publication statusPublished - 17 Sept 1999


  • Actins/metabolism
  • Animals
  • Cell Adhesion
  • Cells, Cultured
  • Cytoskeleton/metabolism
  • Intercellular Adhesion Molecule-1/metabolism
  • Ligands
  • Lymphocyte Function-Associated Antigen-1/genetics
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Conformation
  • Structure-Activity Relationship

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