Abstract
The translation restoring factor (RF) and the eukaryotic initiation factor 2 (eIF-2) stimulating protein (ESP) were purified to near homogeneity from the postribosomal supernatant and the ribosomal salt wash, respectively, of rabbit reticulocyte lysate. They were isolated in the form of eIF-2 complexes, apparently in a 1:1 ratio. Their virtually identical sodium dodecyl sulfate/polyacrylamide gel electrophoretic patterns show, in addition to the eIF-2 .alpha.(38,000), .beta.(52,000) and .gamma.(54,000) bands, peptide bands at .apprx. 80, 65, 57, 40 and 32 kilodaltons. The apparent MW of either complex is .apprx. 450,000, whereas that of free translation restoring factor (RF) is .apprx. 250,000. At 0.5 mM Mg2+, both ESP and RF stimulate ternary complex (eIF-2.cntdot.GTP.cntdot.Met-tRNAi) formation catalytically with unphosphorylated eIF-2. Phosphorylation of the eIF-2 .alpha. subunit by preincubation with eIF-2 .alpha. kinase and ATP, which virtually blocks eIF-2-ESP interaction, results in only partial blocking of the interaction with RF. This may explain the translation-restoring activity of RF.

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