Expression of distinct fucosylated oligosaccharides and carbohydrate-mediated adhesion efficiency directed by two different alpha-1,3-fucosyltransferases. Comparison of E- and L-selectin-mediated adhesion.
Among five different human alpha 1 --> 3 fucosyltransferases cloned, fucosyltransferases III (Fuc-TIII) and IV (Fuc-TIV) differ significantly from each other. Fuc-TIII transfers a fucose to both sialylated and nonsialylated N-acetyllactosamine, but Fuc-TIV apparently transfers a fucose only to neutral N-acetyllactosamine. In this study, Chinese hamster ovary (CHO) cells were stably transfected with Fuc-TIII or Fuc-TIV, and the resultant cell lines, CHO-FTIII and CHO-FTIV, were compared for the carbohydrate structures and for their binding to E-selectin or L-selectin. CHO-FTIII and CHO-FTIV cells were labeled metabolically with [3H]galactose, and glycopeptides obtained from these cells were fractionated by serial lectin affinity chromatography. The fractionated glycopeptides were then subjected to various combinations of exoglycosidase treatment or endo-beta-galactosidase digestion. The results obtained can be summarized as follows. CHO-FTIII cells express sialyl Lewisx, Lewisx, and VIM-2 structures, whereas CHO-FTIV cells express only an Lex structure with a small amount of VIM-2 structure. When CHO-FTIII and CHO-FTIV cells were tested for adhesion to E-selectin expressed by tumor necrosis factor-activated endothelial cells and to an E-selectin chimeric protein, only CHO-FTIII cells were found to adhere well to E-selectin. Moreover, both CHO-FTIII and CHO-FTIV cells failed to adhere to an L-selectin chimeric protein. These results clearly indicate that FT-III and FT-IV direct distinctly different fucosylated oligosaccharides. This difference in oligosaccharide structures results in an entirely different efficiency in adhesion to E-selectin. The results also demonstrate that expression of sialyl Lex itself is not sufficient for L-selectin binding.
Sueyoshi
S
S
Glycobiology Program, La Jolla Cancer Research Foundation, California 92037.
Tsuboi
S
S
Sawada-Hirai
R
R
Dang
U N
UN
Lowe
J B
JB
Fukuda
M
M
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D050260:Carbohydrate Metabolism; D002240:Carbohydrate Sequence; D002448:Cell Adhesion; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002846:Chromatography, Affinity; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D019040:E-Selectin; D005643:Fucose / Q000378:metabolism*; D005647:Fucosyltransferases / Q000378:metabolism*; D006020:Glycopeptides / Q000302:isolation & purification; D019041:L-Selectin; D037102:Lectins; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism
D015815:Cell Adhesion Molecules; D017931:DNA Primers; D019040:E-Selectin; D006020:Glycopeptides; D037102:Lectins; D009844:Oligosaccharides; D011134:Polysaccharides; D019041:L-Selectin; D005643:Fucose; C037199:poly-N-acetyllactosamine; D005647:Fucosyltransferases; C031692:galactoside 3-fucosyltransferase