CEACAM1, a cell-cell adhesion molecule, directly associates with annexin II in a three-dimensional model of mammary morphogenesis

J Biol Chem. 2003 Dec 12;278(50):50338-45. doi: 10.1074/jbc.M309115200. Epub 2003 Sep 30.


The epithelial cell adhesion molecule CEACAM1 (carcinoembryonic antigen cell adhesion molecule-1) is down-regulated in colon, prostate, breast, and liver cancer. Here we show that CEACAM1-4S, a splice form with four Ig-like ectodomains and a short cytoplasmic domain (14 amino acids), directly associates with annexin II, a lipid raft-associated molecule, which is also down-regulated in many cancers. Annexin II was identified using a glutathione S-transferase pull-down assay in which the cytoplasmic domain of CEACAM-4S was fused to glutathione S-transferase, the fusion protein was incubated with cell lysates, and isolated proteins were sequenced by mass spectrometry. The interaction was confirmed first by reciprocal immunoprecipitations using anti-CEACAM1 and anti-annexin II antibodies and second by confocal laser microscopy showing co-localization of CEACAM1 with annexin II in mammary epithelial cells grown in Matrigel. In addition, CEACAM1 co-localized with p11, a component of the tetrameric AIIt complex at the plasma membrane, and with annexin II in secretory vesicles. Immobilized, oriented peptides from the cytoplasmic domain of CEACAM1-4S were shown to directly associate with bovine AIIt, which is 98% homologous to human AIIt, with average KD values of about 30 nM using surface plasmon resonance, demonstrating direct binding of functionally relevant AIIt to the cytoplasmic domain of CEACAM1-4S.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Annexin A2 / chemistry*
  • Annexin A2 / metabolism
  • Antigens, CD / metabolism
  • Antigens, CD / physiology*
  • Antigens, Differentiation / metabolism
  • Antigens, Differentiation / physiology*
  • Blotting, Western
  • Cattle
  • Cell Adhesion
  • Cell Adhesion Molecules
  • Cell Line
  • Cell Line, Tumor
  • Collagen / pharmacology
  • Cytoplasm / metabolism
  • Down-Regulation
  • Drug Combinations
  • Glutathione Transferase / metabolism
  • HeLa Cells
  • Humans
  • Kinetics
  • Laminin / pharmacology
  • Mass Spectrometry
  • Microscopy, Confocal
  • Molecular Sequence Data
  • Peptides / chemistry
  • Precipitin Tests
  • Protein Binding
  • Protein Structure, Tertiary
  • Proteoglycans / pharmacology
  • Recombinant Fusion Proteins / metabolism
  • Surface Plasmon Resonance
  • Time Factors


  • Annexin A2
  • Antigens, CD
  • Antigens, Differentiation
  • CD66 antigens
  • Cell Adhesion Molecules
  • Drug Combinations
  • Laminin
  • Peptides
  • Proteoglycans
  • Recombinant Fusion Proteins
  • matrigel
  • Collagen
  • Glutathione Transferase