Structural determination of two N‐linked glycans isolated from recombinant human lactoferrin expressed in BHK cells

FEBS Letters

Plancke, Yves¹  Salmon, Valérie¹  Benaissa, Monique¹  Spik, Geneviéve¹  Legrand, Dominique¹  Coddeville, Bernadette¹ 

[1] Laboratoire de Chimie Biologique, UMR CNRS n 111, Université des Sciences et Technologies de Lille, 59655 Villeneuve d'Ascq Cedex, France

关键词: Human lactoferrin;    Lactotransferrin;    N-glycan;    Recombinant glycoprotein;    BHK cell;    hLf;    human lactoferrin;    DMEM;    Dulbecco's modified essential medium;    BHK;    baby hamster kidney;    EPO;    erythropoietin;    MTX;    methotrexate;    NMR;    nuclear magnetic resonance;    TLC;    thinlayer chromatography;    GLC;    gas liquid chromatography;   

DOI  :  10.1016/0014-5793(95)00441-B

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 α-2,3-disialylated forms (80%) or α-2,3-monosialylated (20%) forms. Moreover, 70% of total glycans were α-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.

【 授权许可】

Unknown