Dear all, I am working on a protein where I have to stabilize the closed conformation of the protein using disulphide bond. The strategy to design the cysteine mutants is based on the molecular dynamic simulations, and accordingly the residues were chosen. The ultimate goal is to trap the ligand in closed conformation of protein and crystallize it. I am facing few issues: Is there some reliable assay that can check the formation of disulphide bonds in protein. Additionally, does anybody knows another method(s) that can be used to trap a closed conformation. I look forward for your suggestions and discussions on this issue.
Thanks very much! Jan
