Fig. 5.

(A) SDS–PAGE of (1) galectin-1-hum-MFng and (2) hum-MFng after Tev protease cleavage of the fusion protein and purification on a UDP-agarose column. (B) The catalytic activity of the hum-MFng with 500 µM UDP-GlcNAc as the sugar donor substrate and fucose (continuous line) or pNP-fucose (dashed line) as acceptor substrates. Each assay was carried out for 1 h at 37°C. (C) The chemoenzymatic detection of the transferred C2-keto-Glc on O-fucosylated EGF repeat from Factor VII (a kind gift from Dr. R. Haltiwanger). Reaction samples without enzyme (a) and with enzyme (b).