Comparative study of the asparagine-linked sugar chains of human lipocalin-type prostaglandin D synthase purified from urine and amniotic fluid, and recombinantly expressed in Chinese hamster ovary cells.
Lipocalin-type prostaglandin D synthase (L-PGDS) is a highly glycosylated member of the lipocalin gene family and is secreted into various human body fluids. We comparatively analyzed the structures of asparagine-linked sugar chains of human L-PGDS produced by recombinant Chinese hamster ovary cells and naturally occurring human urine and amniotic fluid. After the sugar chains were liberated by hydrazinolysis followed by N-acetylation, they were derivatized with 2-aminobenzamide. All of the sugar chains of three L-PGDSs occur as biantennary complex-type sugar chains. Most of the sugar chains of three samples were fucosylated on the inner most N-acetylglucosamine residue. Although the sugar chains of the recombinant L-PGDS do not contain any bisecting N-acetylglucosamine residues, 58% and 34% of the fucosylated-sugar chains of amniotic fluid and urine L-PGDSs, respectively, contain bisecting N-acetylglucosamine residues. The sialic acid residues occur solely as Siaalpha2-->3Gal groups of the recombinant L-PGDS; the sialic acid residues of other L-PGDS occur as both Siaalpha2-->3Gal and Siaalpha2-->6Gal groups. Variations in L-PGDS glycosylation may prove useful as markers to further elucidate the role of L-PGDS glycoforms in different tissues.
Manya
H
H
Department of Glycobiology, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo 173-0015, Japan.
Sato
Y
Y
Eguchi
N
N
Seiki
K
K
Oda
H
H
Nakajima
H
H
Urade
Y
Y
Endo
T
T
D000117:Acetylglucosamine / Q000032:analysis; D000653:Amniotic Fluid / Q000201:enzymology; D000818:Animals; D001216:Asparagine / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000302:isolation & purification*; D016466:CHO Cells; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D005643:Fucose / Q000032:analysis*; D006031:Glycosylation; D006801:Humans; D019746:Intramolecular Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification* / Q000652:urine; D054834:Lipocalins; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000032:analysis; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D062367:ortho-Aminobenzoates / Q000737:chemistry
D054834:Lipocalins; D009844:Oligosaccharides; D011994:Recombinant Proteins; C086316:glucosylasparagine; D062367:ortho-Aminobenzoates; D005643:Fucose; D001216:Asparagine; D019746:Intramolecular Oxidoreductases; C022466:prostaglandin R2 D-isomerase; D019158:N-Acetylneuraminic Acid; C000219:anthranilamide; D000117:Acetylglucosamine