| Thermal Denaturation Assay | |||
| Instrument | Prometheus NT48 (Nano Temper) | ||
| Employed software | PR. control (NanoTemper) | ||
| Wavelength (nm) | ratio 350 - 330 | ||
| Temperature gradient (˚C) | 20 - 95 | ||
| Heating rate (˚C min−1) | 1 | ||
| Protein sample | PBP4* | PBP4* ΔCTD | |
| Concentration (µM) | 30 | 30 | |
| Buffer condition | 25 mM HEPES, 100 mM NaCl, pH 8.0 | ||
| Volume (µL) | 10 | 10 | |
| Results | |||
| Fusion temperature Tm (˚C) | 59˚C ± 0.3˚C | 60˚C ± 0.2˚C | |
| Dynamic Light Scattering Assay | |||
| Instrument | DynaPro NanoStarTM (Wyatt Technologies) | ||
| Employed software | Dynamics 7.1.9. (Wyatt Technology) | ||
| Laser | 100 mW He-Ne | ||
| Wavelength (nm) | 658 | ||
| Angle (˚) | 90 | ||
| Exposure time (sec) | 50 (10 frames × 5 sec) | ||
| Protein sample | PBP4* | PBP4* ΔCTD | |
| Concentration (µM) | 30 | 30 | |
| Buffer condition | 25 mM HEPES, 100 mM NaCl, pH 8.0 | ||
| Temperature (˚C) | 10 | 10 | |
| Volume (µL) | 50 | 50 | |
| Results | |||
| Hydrodynamic diameter (nm) | 6.57 ± 0.062 | 5.462 ± 0.084 | |
| Polydispersity index (PDI) | 0.016 ± 0.004 | 0.009 ± 0.002 | |
| Estimated molecular weight (kDa) | 54.5 ± 1.22 | 35.17 ± 1.17 | |
| Exclusion Chromatography | |||
| Column | Superdex S200 10/30 GL (GE Healthcare) | ||
| Instrument | Åkta Purifier (GE Healthcare) | ||
| Employed software | Unicorn (GE Healthcare Life Sciences) | ||
| Wavelength (nm) | 280 | ||
| Flow (mL min−1) | 0.5 | ||
| Injection loop volume (µL) | 100 | ||
| Protein sample | PBP4* | PBP4* ΔCTD | |
| Injected mass (µg) | 100 | 100 | |