Determination of dissociation constant of SMT3-A1(Cys) for SMT3 (yeast SUMO) by microscale thermophoresis


Overview
Abstract
The dissociation constant (Kd) of the interaction between SMT3-A1(Cys) and SMT3 (yeast SUMO) was measured by microscale thermophoresis. Note: SMT3-A1 with no Cys was used to eliminate disulfide interactions. SMT3-GFP was held at 10 nM while the concentration of SMT3-A1 was varied from 5 µM to 0.153 nM while the migration of the fluorescent protein was measured upon local heating using a Monolith NT.115 Pico with 20% Laser Power, 15% LED power, at 25C.
Type
  • Biophysical Constant Measurement
Target
Scores
Outcome
Successful
Signal
Acceptable
Results
Figure

Image 

MST determination of dissociation constant for SUMO and nanoCLAMP SMT3-A1

Legend 

Analysis of the interaction between SMT3-A1 and SMT3-GFP (SUMO-GFP) by microscale thermophoresis. Binding of SMT3-A1 to SMT3-GFP fusion protein was quantified in PBS-T (0.05% Tween). SMT3-GFP was held at 10 nM while the concentration of SMT3-A1 was varied from 5 µM to 0.153 nM while the migration of the fluorescent protein was measured upon local heating using a Monolith NT.115 Pico with 20% Laser Power, 15% LED power, at 25C.