6'-Sulfo sialyl Lex but not 6-sulfo sialyl Lex expressed on the cell surface supports L-selectin-mediated adhesion.
In order to determine if a sulfated oligosaccharide on the cell surface can function as an L-selectin ligand, a novel approach for in vitro transfer of oligosaccharides was utilized (Srivastava, G., Kaun, K. J., Hindsgaul, O., and Palcic, M. M. (1992) J. Biol. Chem. 267, 22356-22361). CHO cells were incubated with synthetic 6'-sulfo sialyl Lex, NeuNAcalpha2-->3(sulfate-6)Galbeta1-->4(Fucalpha1-->3) GlcNAc or 6-sulfo sialyl Lex, NeuNAcalpha2-->3Galbeta1-->4[(Fucalpha1-->3)sulfate--> 6GlcNAc] oligosaccharide linked to C-6 of a fucose residue in GDP-fucose and a milk fucosyltransferase. The resultant CHO cells expressing 6'-sulfo sialyl Lex or 6-sulfo sialyl Lex on their cell surface were tested for adhesion to E-selectin and L-selectin chimeric proteins coated on plates. The results indicate that 6'-sulfo sialyl Lex supports L-selectin-mediated adhesion much better than sialyl Lex similarly tagged on the cell surface. In contrast, 6-sulfo sialyl Lex containing a sulfate group on the N-acetylglucosamine residue did not support adhesion with either selectin. These combined results suggest that 6'-sulfo sialyl Lex is a much better ligand than sialyl Lex oligosaccharide for L-selectin.
Tsuboi
S
S
Glycobiology Program, La Jolla Cancer Research Center, The Burnham Institute, La Jolla, California 92037, USA.
Isogai
Y
Y
Hada
N
N
King
J K
JK
Hindsgaul
O
O
Fukuda
M
M
D000818:Animals; D016466:CHO Cells; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002448:Cell Adhesion; D002462:Cell Membrane / Q000502:physiology; D006224:Cricetinae; D005647:Fucosyltransferases / Q000378:metabolism; D006154:Guanosine Diphosphate Fucose / Q000378:metabolism; D019041:L-Selectin / Q000378:metabolism*; D016256:Lewis X Antigen / Q000031:analogs & derivatives; D008892:Milk / Q000201:enzymology; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D000080506:Sialyl Lewis X Antigen / Q000031:analogs & derivatives; D014162:Transfection
C086949:6'-sulfated sialyl Lewis x; D016256:Lewis X Antigen; D009844:Oligosaccharides; D011993:Recombinant Fusion Proteins; D000080506:Sialyl Lewis X Antigen; D019041:L-Selectin; D006154:Guanosine Diphosphate Fucose; D005647:Fucosyltransferases