Formation of Monofluorinated Radical Cofactor in Galactose Oxidase through Copper-Mediated C-F Bond Scission.

ABSTRACT: Galactose oxidase (GAO) contains a Cu(II)-ligand radical cofactor. The cofactor, which is autocatalytically generated through the oxidation of the copper, consists of a cysteine-tyrosine radical (Cys-Tyr^•) as a copper ligand. The formation of the cross-linked thioether bond is accompanied by a C-H bond scission on Tyr272 with few details known thus far. Here, we report the genetic incorporation of 3,5-dichlorotyrosine (Cl₂-Tyr) and 3,5-difluorotyrosine (F₂-Tyr) to replace Tyr272 in the GAO^V previously optimized for expression through directed evolution. The proteins with an unnatural tyrosine residue are catalytically competent. We determined the high-resolution crystal structures of the GAO^V, Cl₂-Tyr272, and F₂-T

SUBMITTER: Li J 

PROVIDER: S-EPMC7737484 | biostudies-literature | 2020 Nov

REPOSITORIES: biostudies-literature