Thermal protein denaturation to validate small molecule-protein interaction
The fluorescence intensity of sypro orange increases significantly when it binds to exposed hydrophobic regions on proteins. This has been used to monitor the thermal denaturation of proteins. PCL is currently optimizing the thermal melting of carbonic anhydrase using the CFX96 Real-Time PCR instrument by monitoring the change in sypro orange intensity. Once this phase of the work is complete, PCL will monitor the effect of known small molecule binders on the thermal denaturation of carbonic anhydrase.