The binding specificity of normal and variant rat Kupffer cell (lectin) receptors expressed in COS cells.
A receptor uniquely found on the surface of rat Kupffer cells was shown previously to bind oligosaccharides terminating in galactose, N-acetylgalactosamine, and fucose. To analyze further the binding specificity of the receptor, receptor-mediated adhesion of transfected COS cells to immobilized glycolipids of known structure was measured. The glycolipid Gb4Cer (GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1Cer) was the best ligand. Gb5Cer (GalNAc alpha 1-3GalNAc beta 1-3Gal alpha 1-4Gal beta 1-4Glc beta 1Cer) and LacCer (Gal beta 1-4Glc beta 1Cer) bound more weakly (five times less than Gb4Cer) and Gb3Cer (Gal alpha 1-4Gal beta 1-4Glc beta 1Cer), and g3Cer(GalNAc beta 1-4Gal beta 1-4Glc beta 1Cer) bound even more weakly (60 times less than Gb4Cer). Gangliosides did not support adhesion of transfected cells. The adhesion of COS cells transfected with plasmids encoding variants of the receptor was also examined. In each variant, either tryptophan 498 or 523, which are conserved in most C-type lectins, was replaced by one of several amino acids. Variants that retained binding activity had the same specificity as the normal receptor. Differences between variants were noted, however, in maximal levels of adhesion and these differences correlated with altered expression of the receptor variants in COS cells.
Tiemeyer
M
M
Glycomed Incorporated, Alameda, California 94501.
Brandley
B K
BK
Ishihara
M
M
Swiedler
S J
SJ
Greene
J
J
Hoyle
G W
GW
Hill
R L
RL
D000116:Acetylgalactosamine / Q000378:metabolism; D000818:Animals; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D002460:Cell Line; D002855:Chromatography, Thin Layer; D005643:Fucose / Q000378:metabolism; D005690:Galactose / Q000378:metabolism; D007728:Kupffer Cells / Q000378:metabolism*; D008969:Molecular Sequence Data; D010957:Plasmids; D051381:Rats; D011975:Receptors, Mitogen / Q000378:metabolism*; D013379:Substrate Specificity; D014162:Transfection
D011975:Receptors, Mitogen; D005643:Fucose; D000116:Acetylgalactosamine; D005690:Galactose