METHOD FOR INCORPORATING INTERNAL POLAR AND IONIZABLE GROUPS IN PROTEINS

摘要

Internal polar and ionizable groups are essential for enzymatic catalysis, proton transport, redox reactions, and many other functional properties of proteins. To engineer novel enzymes or to modify the function of existing ones, and to build switches that can be used to modify the stability of proteins in response to changes in pH, it is necessary to introduce polar or ionizable groups or to modify the properties of existing ones. However, internal polar and ionizable groups usually destabilize proteins. The disclosure provides new methods that allow the introduction of polar and ionizable groups into the interior of proteins, as well as new methods for improving the accuracy of pKa of an internal amino acid of a protein, and methods for mapping the folding free energy landscape of a protein.