Abstract

The p-aminophenyl gamma-ester of 7-methylguanosine 5'-triphosphate (m7GTP) was synthesized and coupled to Sepharose 4B. A 0.5 M salt extract of rabbit reticulocyte ribosomes was passed over a column containing the affinity medium. After extensive washing, a solution of m7GTP was passed through the column, and a single polypeptide species of 24 kilodaltons (kDa) was eluted. This had an electrophoretic mobility identical with that of the mRNA cap-binding protein. This assignment was confirmed by the fact that the eluted material was enriched nearly 200-fold in the ability to specifically bind 32P-labeled capped oligonucleotides. A control affinity medium consisting of GTP similarly coupled to Sepharose failed to retain the 24-kDa species. The postribosomal supernatant fraction yielded slightly more of the 24-kDa species than the ribosomal wash fraction when passed over this affinity medium.