H (0) blood group determinant is present on soluble human L-selectin expressed in BHK-cells.
In the present study we show that the H (0) blood group determinant Fuc alpha1-2Gal beta1-4GlcNAc beta1-R is present on N-linked glycans of soluble human L-selectin recombinantly expressed in baby hamster kidney (BHK) cells. The glycans were isolated using complementary HPLC techniques and characterized by a combination of exoglycosidase digestion and mass spectrometry. The linkage of the fucose residues was determined by incubation of the glycans with specific fucosidases. The H blood determinant Fuc alpha1-2Gal beta1-4GlcNAc beta1 was detected for bi-, 2,4 branched tri- and tetraantennary structures. To our knowledge, the proposed oligosaccharide structures represent a new glycosylation motif for recombinant glycoproteins expressed on BHK cells.
Gohlke
M
M
Institut für Molekularbiologie und Biochemie der Freien Universität Berlin, Berlin-Dahlem, Germany. rolfnuck@zedat.fu-berlin.de
Mach
U
U
Nuck
R
R
Volz
B
B
Fieger
C
C
Tauber
R
R
Reutter
W
W
D000017:ABO Blood-Group System / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D006224:Cricetinae; D000939:Epitopes / Q000032:analysis*; D006023:Glycoproteins / Q000737:chemistry; D006801:Humans; D007231:Infant, Newborn; D007668:Kidney / Q000378:metabolism; D019041:L-Selectin / Q000737:chemistry*; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry / Q000276:immunology; D017421:Sequence Analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000017:ABO Blood-Group System; D000939:Epitopes; D006023:Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; D019041:L-Selectin