The Free Radical in Ribonucleotide Reductase from E. coli

Abstract

Protein B2, one of the subunits of ribonucleotide reductase from Escherichia coli, contains a stable free radical. It is characterized by a doublet e.p.r. signal centered around g = 2.0047 and a sharp peak at 410 nm in the optical spectrum. The radical has been assigned to a tyrosyl residue in the protein with its spin density delocalized over the aromatic ring. Protein B2 also contains two antiferromagnetically-coupled high-spin iron(III) atoms, which stabilize the free radical. Protein B1, the other subunit of ribonucleotide reductase, contains two binding sites for substrate molecules, which are the four common ribonucleoside diphosphates. It also contains two classes of allosteric effector-binding sites. ATP and deoxyribonucleoside triphosphates function as effectors. A one-to-one complex of proteins B1 and B2 forms the enzymically-active ribonucleotide reductase. The free radical is, most likely, part of the active site.