Rat liver ribonucleotide reductase: separation, purification, and properties of two nonidentical subunits

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Abstract
Two nonidentical subunits of mammalian ribonucleotide reductase, L1 and L2, from regenerating rat liver were extensively purified for the first time. They were separated by dATP-Sepharose affinity chromatography. Subunit L, which bound to dATP-Sepharose, was eluted with 50 mM ATP and purified to homogeneity (as demonstrated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis) by molecular exclusion high-pressure liquid chromatography (HPLC). This subunit had an apparent relative mass (mr) of 45,000 and a Km of 0.9 .times. 10-4 for CDP. Subunit L2, which did not bind to dATP-Sepharose, was purified by pH 5.2 precipitation followed by chromatography on CM-Sephadex, molecular exclusion HPLC and DEAE-cellulose. This subunit contained Fe and had an apparent (mr) of 120,000 by HPLC molecular exclusion chromatography, but showed 2 bands (mr 75,000 and mr) 47,000) on SDS-polyacrylamide gel electrophoresis. Neither L1 nor L2 separately had any enzyme activity, but when combined they reduced CDP to dCDP.

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