Table 1.
Physical properties of the studied proteins.
| Protein | Mass (Da) | pI (Det.) | pI (Calc.) | Tm (°C) | Rate of sterol transfer (%) |
|---|---|---|---|---|---|
| β-Cryptogein | 10385 | 9.8 | 9.7 | 60.59 ± 0.03 | 100 |
| K39T | 10357 | 9.4 | 9.5 | 61.46 ± 0.19 | 67 |
| K48T | 10356 | 9.7 | 9.5 | 61.84 ± 0.08 | 61 |
| K94T | 10356 | 10 | 9.5 | 50.20 ± 1.04 | 135 |
| K39T/K94T | 10328 | 8.7 | 9.1 | ND | 65 |
| K61T/K62T | 10329 | 7.1 | 9.1 | ND | 59 |
| K39T/K48T/K94T | 10301 | 6.7 | 6.7 | ND | 66 |
The molecular mass of the proteins was determined by MALDI-TOF analysis. pI values were calculated by EMBOSS explorer (http://emboss.bioinformatics.nl) with three disulfide bridges or were measured by the IEF method on IPG strips (3–10). Tm values, which reflect the thermodynamic stability of the studied protein, were measured by the protein thermal shift assay (TSA) using the SYPRO Orange fluorescent dye. The rate of sterol transfer was measured from the increase in fluorescence after 2 min due to dilution of DHE in the stigmasterol micelles. ND, not detected.