Table 1: Kinetic parameters of human FUT8 using GDP-Fucose as the donor substrate and different N-glycans as the acceptor substrates.
The kinetic parameters of enzyme activity was obtained via the UDP-Glo assays. The concentration range of the glycan substrates was 0–1.5 mM for the Man5GlcNAc2-Asn substrate and 0–1 mM for others.
| Acceptor Substrates | kcat | KM | kcat/KM |
|---|---|---|---|
| s−1 | μM | mM−1s−1 | |
| Man5GlcNAc2-Asn | 0.052 ± 0.005 | 2114 ± 291 | 0.025 ± 0.006 |
| Man5GlcNAc2-Asn-Fmoc | 0.214 ± 0.045 | 1151 ± 384 | 0.186 ± 0.101 |
| GlcNAc1Man5GlcNAc2Asn | 8.5 ± 0.4 | 234.9 ± 24.5 | 36.1 ± 5.3 |
| GlcNAc1Man5GlcNAc2Asn -Fmoc | 11.6 ± 0.6 | 30.6 ± 7.4 | 378.7 ± 110.4 |
| GlcNAc2Man3GlcNAc2Asn (A2-Asn) | 11.8 ± 0.4 | 52.1 ± 6.6 | 226.4 ± 36.3 |
| GlcNAc2Man3GlcNAc2Asn-Fmoc (A2-Asn-Fmoc) | 13.6 ± 0.5 | 44.9 ± 6.5 | 302.3 ± 55.4 |