Table 1.
Binding affinity of EndoS2 for IgG antibodies and N-glycans
| N° | Enzyme | Ligand | KD (µM) | koff (s−1) | kon (M−1s−1) |
|---|---|---|---|---|---|
| 1 | EndoS2E186L | Fc | 3.1 ± 0.6 | 4.0 ± 1.5 | 1.3 × 106 ± 5.5 × 105 |
| 2 | EndoBT-3987D312A/E314L | HM | 1.7 ± 0.9 | 5.5 ± 1.4 | 3.2 × 106 ± 1.9 × 105 |
| 3 | EndoS2E186L | CT | 106.9 ± 6.7 | 11.4 ± 1.5 | 1.1 × 105 ± 1.6 × 104 |
| 4 | EndoS2E186L | CTn−1 | 150.7 ± 18.0 | 9.4 ± 3.7 | 6.2 × 104 ± 2.6 × 104 |
| 5 | EndoS2E186L | Fcaglyc | – | – | – |
| 6 | EndoS2E186L | CaCl2 | 3415 ± 480 | – | – |
| 7 | EndoS2 | CaCl2 | 4000 ± 100 | – | – |
Dissociation constants KD, on- and off-rate constants kon and koff for Rituximab-Fc, CT, CTn-1 HM and Ca2+ binding to EndoS2E186L, EndoS2 and EndoBT-3987D312A/E314L from fitting a two-state binding model to methyl-TROSY titration data using TITAN algorithm.