Dictyostelium cytosolic fucosyltransferase synthesizes H type 1 trisaccharide in vitro

FEBS Letters

Trinchera, Marco¹  Bozzaro, Salvatore² 

[1] Department of Biochemistry, University of Pavia, via Taramelli 3B, 27100 Pavia, Italy;Department of Clinical and Biological Sciences, University of Torino, Ospedale S. Luigi, 10043 Orbassano (TO), Italy

关键词: Glycosyltransferase;    Cytosol;    Carbohydrate antigen;    Permethylation;    (Dictyostelium discoideum);    Lewis a trisaccharide;    Galβ1-3[Fucα1-4]GlcNAc;    H type 1 trisaccharide;    Fucα1-2Galβ1-3GlcNAc;    lacto-N-biose I;    Galβ1-3GlcNAc;    LacNAc;    Galβ1-4GlcNAc;    GDP-Fuc;    GDP-β-l-fucose;    NEM;    N-ethylmaleimide;    TLCK;    chloromethyl ketone;    PMSF;    phenylmethylsulfonyl fluoride;    HPTLC;    high-performance thin-layer chromatography;    DMSO;    dimethylsulfoxide;    Dα1;    2FucT;    Dictyostelium cytosolic α1;    2fucosyltransferase;    Hα1;    2FucT;    human blood group H α1;    2fucosyltransferase;   

DOI  :  10.1016/0014-5793(96)01003-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A fucosyltransferase activity has been detected using lacto-N-biose I as acceptor in the lower eukaryote Dictyostelium discoideum. This transferase requires divalent cations and is inhibited by N-ethylmaleimide and detergent treatment. Apparent calculated K _m values for GDP-Fuc and lacto-N-biose I are 1.27 μM and 2.80 mM, respectively. The activity is quantitatively recovered in the supernatant after centrifugation at 100 000 × g for 1 h. The reaction product, as determined by gel permeation chromatography, sensitivity to fucosidases, and analysis of partially methylated derivatives, is Fucα1-2Galβ1-3GlcNAc (H type 1 trisaccharide).

【 授权许可】

Unknown