Recognition of diverse proteins by members of the immunoglobulin superfamily: delineation of the receptor binding site in the human CD6 ligand ALCAM.
The CD6-ALCAM (activated leukocyte cell adhesion molecule) interaction, which mediates thymocyte--thymic epithelial cell adhesion, is a previously unobserved type of protein--protein interaction that involves members of the scavenger receptor cysteine rich protein superfamily (SRCRSF) and the immunoglobulin superfamily (IgSF). Targeted mutagenesis of ALCAM reveals that residues which constitute the CD6 binding site cluster on the predicted A'GFCC'C" face of its N-terminal Ig domain. These results, in conjunction with recent analyses of interactions involving other IgSF members, suggest that this region in IgSF cell surface proteins is most suitable to mediate interactions with different ligands irrespective of their structure. The CD6 binding site in ALCAM is conserved across species, and nonconserved residues in ALCAM and its murine homolog map to the beta-sheet face opposite to the CD6 binding site. This provides a molecular rationale for the inability to obtain murine monoclonal antibodies against the receptor binding domain which block the CD6-ALCAM interaction.
Skonier
J E
JE
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121, USA.
Bowen
M A
MA
Emswiler
J
J
Aruffo
A
A
Bajorath
J
J
D020410:Activated-Leukocyte Cell Adhesion Molecule; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015703:Antigens, CD / Q000378:metabolism*; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism*; D001665:Binding Sites; D015815:Cell Adhesion Molecules / Q000737:chemistry* / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D017124:Conserved Sequence / Q000235:genetics; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017385:Sequence Homology; D013601:T-Lymphocytes / Q000737:chemistry / Q000378:metabolism
D020410:Activated-Leukocyte Cell Adhesion Molecule; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; C062279:CD6 antigen; D015815:Cell Adhesion Molecules; D006023:Glycoproteins; D007136:Immunoglobulins