Association of the cytoplasmic domain of intercellular-adhesion molecule-1 with glyceraldehyde-3-phosphate dehydrogenase and beta-tubulin

Eur J Biochem. 1996 May 15;238(1):173-80. doi: 10.1111/j.1432-1033.1996.0173q.x.


To elucidate the molecular mechanisms of the transendothelial migration of leukocytes, we attempted to identify the cellular proteins capable of interaction with the cytoplasmic domain of the intercellular adhesion molecule-1 (ICAM-1) in a rat brain microvessel endothelial cell line (RBE4 cells). A 27-amino-acid synthetic peptide, corresponding to the cytoplasmic domain of rat ICAM-1, was covalently linked to a Sepharose matrix. Upon affinity chromatography of RBE4 cell cytosol, several ICAM-1-interacting proteins were specifically eluted by the soluble peptide. Two of these proteins have been identified by microsequencing as the cytoskeletal protein beta-tubulin and the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GraP-DH). Experiments carried out with purified GraP-DH or CNBr fragments of GraP-DH indicated that binding to the ICAM-1 matrix was mediated by the C-terminal domain of GraP-DH, containing the binding site of the cofactor NAD+, and that NAD+ could compete with this binding. Using a series of ICAM-1 C-terminal truncated peptides, we could demonstrate that (a) the nitric-oxide-induced covalent linkage of NAD+ to GraP-DH was impaired by these peptides, (b) the glycolytic activity of GraP-DH was drastically inhibited by a truncated peptide containing the 15 C-terminal residues, (c) nitric oxide appeared to prevent this inhibition. Together, our results demonstrate that GraP-DH specifically associates with the isolated ICAM-1 cytoplasmic domain. Since GraP-DH is known as a microtubule bundling protein, these findings suggest that, in a cellular environment, GraP-DH may behave as an adaptor molecule by linking ICAM-1 to the microtubule network. The role of nitric oxide in the modulation of this interaction deserves further investigation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Binding, Competitive
  • Brain / metabolism
  • Cytoplasm / metabolism*
  • Cytosol / chemistry
  • Cytosol / metabolism
  • Glyceraldehyde-3-Phosphate Dehydrogenases / drug effects
  • Glyceraldehyde-3-Phosphate Dehydrogenases / metabolism*
  • Intercellular Adhesion Molecule-1 / metabolism*
  • Molecular Sequence Data
  • NAD / drug effects
  • NAD / metabolism
  • Nitric Oxide / metabolism
  • Nitric Oxide / pharmacology
  • Peptide Fragments / metabolism
  • Peptide Fragments / pharmacology
  • Rats
  • Signal Transduction
  • Substrate Specificity
  • Tubulin / metabolism*


  • Peptide Fragments
  • Tubulin
  • NAD
  • Intercellular Adhesion Molecule-1
  • Nitric Oxide
  • Glyceraldehyde-3-Phosphate Dehydrogenases