MECHANISM OF LACTOBACILLUS-LEICHMANNII RIBONUCLEOTIDE REDUCTASE STUDIED WITH C-OMICRON-ALPHA-[ALPHA-(ADEN-9-YL)]-C-OMICRON-BETA-ADENOSYLCOBAMIDE (PSEUDOCOENZYME-B12) AS COENZYME

COα-[α-(Aden-9-yl)]-Coβ-adenosylcobamide (pseudocoenzyme B12) purified from Clostridium tetanomorphum has been reacted with ribonucleotide reductase purified from Lactobacillus leichmannii under various conditions, and the properties of the products obtained have been compared by electron paramagnetic resonance (EPR) with those previously reported for products formed from the normal coenzyme (adenosylcobalamin). The rapidly formed intermediate and the slowly formed “doublet” species from the pseudocoenzyme have EPR spectra identical with those formed from the normal coenzyme. This and other considerations make it less likely that the unusual magnetic properties of the rapidly formed intermediate are due to strongly distorted octahedral symmetry about Co(II) as previously postulated. Instead it is probable that the EPR spectrum is due to interaction of the radical pair by both exchange coupling and magnetic dipole-dipole coupling. Although Coα-[α-(aden-9-yl)]cob(II)amide in solution does not show superhyperfine splitting in the EPR spectrum because of its base-off configuration, the cob(II)amide formed by degradation of the pseudocoenzyme within the catalytic site of the enzyme did show triplets due to a nitrogen axially coordinated to cobalt. This suggests that binding of the cob(II)amide to the reductase catalytic site causes a shift to the base-on form. © 1979, American Chemical Society. All rights reserved.