Table 3.
Molecular properties of the proteins determined by SEC-MALS
| Protein | Number of N-glyc sites | N-Glyc state | MWTheoretical (kDa)‡ | MWMeasured (KDa) |
|---|---|---|---|---|
| Fz4CRD | 2 | deglyc* | 17.1 (monomer) | 15.7 ± 0.4 |
| Fz4CRD | 2 | glyc† | 21.4 (monomer) | 23.6 ± 0.3 |
| mFz5CRD | 2 | glyc† | 22.2 (monomer) | 23.9 ± 0.9 |
| mFz8CRD | 2 | glyc† | 22.1 (monomer) | 23.7 ± 0.2 |
| Norrin–Fz4CRD | 4 (2:2 complex) | deglyc* | 61.3 (2:2 complex) | 60.1 ± 0.4 |
| Norrin–Fz4CRD | 4 (2:2 complex) | glyc† | 69.9 (2:2 complex) | 61.3 ± 0.5 |
*
The proteins were produced from HEK293T cells in the presence of the N-glycosylation processing inhibitors, kifunensine resulting in limited glycosylation and were treated with endoglycosidase-F1.
†
The proteins were produced from HEK293T cells with full glycosylation.
‡
The measured molecular weight (MWMeasured) is in general agreement with theoretical molecular weight (MWTheoretical) predicated based on the primary sequence plus the molecular weight of N-linked glycans (see ‘Materials and methods’, SEC-MELS analysis for detailed information of calculation).