Using affinity chromatography to engineer and characterize pH-dependent protein switches.

ABSTRACT: Conformational changes play important roles in the regulation of many enzymatic reactions. Specific motions of side chains, secondary structures, or entire protein domains facilitate the precise control of substrate selection, binding, and catalysis. Likewise, the engineering of allostery into proteins is envisioned to enable unprecedented control of chemical reactions and molecular assembly processes. We here study the structural effects of engineered ionizable residues in the core of the glutathione-S-transferase to convert this protein into a pH-dependent allosteric protein. The underlying rational of these substitutions is that in the neutral state, an uncharged residue is compatible with the hydrophobic environment. In the charged state, however, the residue will invoke unfavorable in

SUBMITTER: Sagermann M 

PROVIDER: S-EPMC2708033 | biostudies-literature | 2009 Jan

REPOSITORIES: biostudies-literature