L-14 lectin recognition of laminin and its promotion of in vitro cell adhesion.
The S-type lactose-binding lectins are found in a variety of animal tissues and their primary sequences are highly conserved between species. Little is known, however, about their functions and endogenous ligands. We report here our studies on the carbohydrate binding specificity of one S-type lectin designated L-14, using glycopeptides and glycoproteins from Chinese hamster ovary cells and mouse teratocarcinoma F9 cells. Our studies demonstrate that L-14 binds with high affinity to oligosaccharides containing multiple repeating units (approximately 4) of the disaccharide [3Gal beta 1-4GlcNAc beta 1]n or poly-N-acetyllactosamine (PL) sequence. Interestingly, terminal beta-galactosyl residues are not necessary for high affinity binding of long PL-containing oligosaccharides to L-14. To characterize the glycoprotein ligands for L-14, we applied total [3H]galactose-labeled glycoproteins from differentiated F9 cells to L-14-Sepharose. Laminin was one of the major glycoproteins in both the cells and the media bound by the lectin. The possible functional significance of this interaction is suggested by the fact that in the absence of Ca2+ L-14 can mediate the binding in vitro of both CHO and F9 cells to immobilized laminin. Taken together, these studies demonstrate that L-14 binds with high affinity to laminin and to relatively long PL chains and indicate that L-14 can promote cell adhesion to laminin.
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Department of Biochemistry and Molecular Biology, Oklahoma Center for Molecular Medicine, University of Oklahoma Health Sciences Center, Oklahoma City 73104.
Cummings
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D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002448:Cell Adhesion / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002846:Chromatography, Affinity; D006224:Cricetinae; D003545:Cysteine / Q000378:metabolism; D004187:Disaccharides / Q000032:analysis*; D004797:Enzyme-Linked Immunosorbent Assay; D005690:Galactose / Q000378:metabolism; D005697:Galactosides / Q000032:analysis*; D037161:Galectins; D006020:Glycopeptides / Q000096:biosynthesis / Q000302:isolation & purification; D006023:Glycoproteins / Q000096:biosynthesis / Q000302:isolation & purification; D006388:Hemagglutinins; D007797:Laminin / Q000032:analysis*; D008715:Methionine / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000032:analysis*; D013462:Sulfur Radioisotopes; D013724:Teratoma; D014212:Tretinoin / Q000494:pharmacology; D014316:Tritium; D014407:Tumor Cells, Cultured
D004187:Disaccharides; D005697:Galactosides; D037161:Galectins; D006020:Glycopeptides; D006023:Glycoproteins; D006388:Hemagglutinins; D007797:Laminin; D011134:Polysaccharides; D013462:Sulfur Radioisotopes; D014316:Tritium; D014212:Tretinoin; D003994:Bucladesine; C037199:poly-N-acetyllactosamine; D008715:Methionine; D003545:Cysteine; D005690:Galactose