Structural determination of two N-linked glycans isolated from recombinant human lactoferrin expressed in BHK cells.
A full-length cDNA coding for human lactoferrin was isolated from a mammary gland library and the recombinant protein was expressed in BHK cells as described by Stowell K. M. et al. [1991, Biochem. J. 276, 349-355]. Two N-linked glycans from purified recombinant lactoferrin were released by hydrazinolysis and analyzed by 400-MHz 1H-NMR spectroscopy. The identified structures corresponded to N-acetyllactosaminic biantennary glycans and were alpha-2,3-disialylated forms (80%) or alpha-2,3-monosialylated (20%) forms. Moreover, 70% of total glycans were alpha-1,6-fucosylated at the GlcNAc residue linked to asparagine. In regard to its glycan moiety, the recombinant glycoprotein is close to native lactoferrins from milk or leucocytes but shows specific structural features which should be taken into account prior to in vivo and in vitro biological studies.
Legrand
D
D
Laboratoire de Chimie Biologique, UMR CNRS n.111, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Salmon
V
V
Coddeville
B
B
Benaissa
M
M
Plancke
Y
Y
Spik
G
G
D000818:Animals; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D006224:Cricetinae; D006031:Glycosylation; D006801:Humans; D007781:Lactoferrin / Q000737:chemistry* / Q000235:genetics; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009005:Monosaccharides / Q000032:analysis; D011134:Polysaccharides / Q000737:chemistry* / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry; D017421:Sequence Analysis
D009005:Monosaccharides; D011134:Polysaccharides; D011994:Recombinant Proteins; D007781:Lactoferrin