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. 2019 Jan 31;20(3):622. doi: 10.3390/ijms20030622

Figure 4.

Figure 4

Enzymatic characterization of the ST8Sia VI enzymes using the MicroPlate Sialyltransferase Assay (MPSA). (A) Time course of human and zebrafish ST8Sia activities onto bovine fetuin. Sialylation reactions were conducted at 25 °C for various incubation times (2–16 h) and 100 µM CMP-SiaNAl with either pFLAG-CMV-9/Dre ST8Sia VI-like or mock HEK293-transfected cells (27.5 µL) or 8 µg of recombinant human ST8Sia VI, n = 2. (B) Human and zebrafish ST8Sia activities on various mammalian glycoproteins using the MPSA. Sialylation reactions were conducted for 4 h at 27 °C using potential acceptor substrates (fetuin, asialofetuin, orosomucoid, asialoorosomucoid and bovine submaxillary mucin (BSM) and 100 µM CMP-SiaNAl with variable amounts (3, 10, 27.5 µL) of cell culture media from pFLAG-CMV-9/Dre ST8Sia VI-like or mock HEK293-transfected cells or recombinant human ST8Sia VI (1–8 µg). Data shown on the left side are those corresponding to the human enzyme and those on the right side are those of the zebrafish enzyme, n = 2.