Synthesis of ATP‐ and dATP‐Substituted Sepharoses and Their Application in the Purification of Phage‐T4–Induced Ribonucleotide Reductase

Abstract

p ‐Aminophenylesters of ATP and dATP were synthesized and coupled to Sepharose 4B activated by cyanogen bromide. On chromatography of a crude extract of T4‐infected Escherichia coli cells on ATP‐ or dATP‐Sepharose columns a limited number of proteins was adsorbed. They could subsequently be eluted from such columns with ATP or dATP as revealed by dodecylsulfate‐polyacrylamide gel electrophoresis. T4 ribonucleotide reductase was purified on dATP‐Sepharose to about 70% purity from a crude extract, and essentially to homogeneity from a crude ammonium sulfate fraction.