Colocalization of the homophilic binding site and the neuritogenic activity of the cell adhesion molecule L1 to its second Ig-like domain.
The cell adhesion molecule L1 has been implicated in mediating cell-cell adhesion and in promoting neurite outgrowth. The extracellular region of L1 contains six immunoglobulin (Ig)-like domains in the amino-terminal region, followed by five fibronectin type III-like repeats. L1 is capable of undergoing homophilic binding as well as heterophilic interactions. To map the homophilic binding domain in L1, three glutathione S-transferase (GST) fusion proteins (GST-Ig1-2-3, GST-Ig4-5-6, and GST-Fn) were prepared and coupled to Covaspheres and their homophilic binding activity was determined using the Covasphere-to-substratum binding assay. Only GST-Ig1-2-3 was capable of homophilic binding. Next, His-tagged recombinant Ig-domain proteins (His-Ig1-2, His-Ig1, and His-Ig2) were expressed and subjected to similar assays. Only His-Ig1-2 and His-Ig2 were capable of homophilic interactions. Binding of His-Ig2-conjugated Covaspheres to substrate-coated His-Ig2 was inhibited by anti-Ig1-2-3 Fab and soluble His-Ig2. These results indicate that the L1 homophilic binding site resides within Ig2. To examine effects of these L1 recombinant proteins on neurite outgrowth, neural retinal cells were cultured on different substrate-coated fusion proteins. Both GST-Ig1-2-3 and His-Ig2 were potent inducers of neurite extension. These results thus indicate that the L1 Ig-like domain 2 alone is sufficient to mediate L1-L1 interaction and promote neurite outgrowth from retinal cells.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002642:Chick Embryo; D007136:Immunoglobulins / Q000737:chemistry*; D039841:Leukocyte L1 Antigen Complex; D008969:Molecular Sequence Data; D019006:Neural Cell Adhesion Molecules / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects*; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship
D007136:Immunoglobulins; D039841:Leukocyte L1 Antigen Complex; D019006:Neural Cell Adhesion Molecules; D011993:Recombinant Fusion Proteins