Active site center redesign increases protein stability preserving catalysis in thioredoxin.

ABSTRACT: The stabilization of natural proteins is a long-standing desired goal in protein engineering. Optimizing the hydrophobicity of the protein core often results in extensive stability enhancements. However, the presence of totally or partially buried catalytic charged residues, essential for protein function, has limited the applicability of this strategy. Here, focusing on the thioredoxin, we aimed to augment protein stability by removing buried charged residues in the active site without loss of catalytic activity. To this end, we performed a charged-to-hydrophobic substitution of a buried and functional group, resulting in a significant stability increase yet abolishing catalytic activity. Then, to simulate the catalytic role of the buried ionizable group, we designed a combinatorial libra

SUBMITTER: Romero ML 

PROVIDER: S-EPMC9601870 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature