Figure 2. Domains involved in PtAu1a dimerization.
Normalized MALS detection of PtAu1afull (a), PtAu1abZIP-LOV (b) and PtAu1aLOV (c) fractionated by size-exclusion chromatography in the dark (black traces) and light (blue traces). The MALS-derived molar-mass signals are shown in green (dark runs) and blue–green (light runs). Additional experiments performed for PtAu1afull and PtAu1abZIP-LOV in the light at varying protein concentrations are shown in Figure 2—figure supplement 1. (d) Quantification of the monomer-dimer equilibrium of PtAu1aLOV in the dark by MST. Error bars represent the standard deviation of three individual experiments. MALS, multi-angle light scattering; MST, microscale thermophoresis.
Figure 2—figure supplement 1. Concentration dependent elution profiles of PtAu1afull and PtAu1abZIP-LOV in the light.
Normalized MALS detection of PtAu1afull (a) and PtAu1abZIP-LOV (b) fractionated by size-exclusion chromatography at protein concentrations of 200 µM (black traces) and 100 µM (dashed gray traces) in the light. The MALS-derived molar mass signals are shown in green (200 µM) and yellow–green (100 µM). PtAu1afull and PtAu1abZIP-LOV were pre-incubated at 20°C under continuous blue light illumination (400 μW cm-2 at 450 nm) from a royal blue (455 nm) collimated LED lamp (Thorlabs) for 5 min. 100 μl protein solution was subjected to size-exclusion chromatography at RT on a Superdex 200 Increase 10/300 GL column (GE Healthcare, Uppsala, Sweden) equilibrated in buffer C. LED, light emitting diode; MALS, multi-angle light scattering; RT, room temperature.