Selenocysteine Substitution in a Class I Ribonucleotide Reductase.

ABSTRACT: Ribonucleotide reductases (RNRs) employ a complex radical-based mechanism during nucleotide reduction involving multiple active site cysteines that both activate the substrate and reduce it. Using an engineered allo-tRNA, we substituted two active site cysteines with distinct function in the class Ia RNR of Escherichia coli for selenocysteine (U) via amber codon suppression, with efficiency and selectivity enabling biochemical and biophysical studies. Examination of the interactions of the C₄₃₉U α₂ mutant protein with nucleotide substrates and the cognate β₂ subunit demonstrates that the endogenous Y₁₂₂^• of β₂ is reduced under turnover conditions, presumably through radical transfer to form a transient U₄₃₉

SUBMITTER: Greene BL 

PROVIDER: S-EPMC6994234 | biostudies-literature | 2019 Dec

REPOSITORIES: biostudies-literature