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Papers In Press, published online ahead of print August 23, 2005
Experimental Immunology Branch, NCI, NIH, Bethesda, MD 20892
Corresponding Author: chavakist{at}mail.nih.gov
The junctional adhesion molecule-C (JAM-C) was recently shown to undergo a heterophilic interaction with the leukocyte
J. Biol. Chem, 10.1074/jbc.M505059200
Submitted on May 9, 2005
Accepted on August 23, 2005
The homophilic binding of junctional adhesion molecule-C mediates tumor cell-endothelial cell interactions
2-integrin Mac-1, thereby mediating interactions between vascular cells in inflammatory cell recruitment. Here, the homophilic interaction of JAM-C is presented and functionally characterized to mediate tumor cell-endothelial cell interactions. Recombinant soluble JAM-C in fluid phase bound to immobilized JAM-C as assessed in a purified system; moreover, JAM-C-transfected CHO cells adhered to immobilized JAM-C. The homophilic interaction of JAM-C was mediated by the isolated amino-terminal Ig-domain (D1) but not the carboxy-terminal Ig-domain (D2) of the molecule. Dimerisation of JAM-A is dependent on the sequence RVE in the aminoterminal Ig-domain. This motif is conserved in JAM-C (R64I65E66) and a single amino acid mutation in this motif (E66R) abolished the homophilic interaction of JAM-C. The lung carcinoma cell line NCI-H522 was found to express JAM-C. NCI-H522 cells adhered to immobilized JAM-C, as well as to JAM-C-transfected CHO cells, but not to mock-transfected CHO cells or to CHO cells transfected with the JAM-C mutant (E66R). Adhesion of NCI-H522 cells to JAM-C protein or JAM-C-transfected CHO cells was abolished in the presence of soluble JAM-C or the isolated D1. Furthermore, the adhesion of NCI-H522 cells to endothelial cells was significantly blocked by soluble JAM-C or the isolated D1. Thus, JAM-C undergoes a homophilic interaction via the R64I65E66 motif on the membrane-distal Ig-domain of the molecule. The homophilic interaction of JAM-C can mediate tumor cell-endothelial cell interactions and may thereby be involved in the process of tumor cell metastasis.
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