1. 1. Affinity chromatography onβ,γ-methylene guanosine 5'-triphosphate-Sepharose was used to purify protein synthesis initiation factor eIF-2 from chicken reticulocytes. 2. 2. Gel filtration of the purified factor gave a molecular weight of 150,000, whereas electrophoresis of the purified factor on polyacrylamide gel containing sodium dodecyl sulfate revealed three non-identical subunits with apparent molecular weight of 57,000, 41,000 and 33,000. 3. 3. With Met-tRNA and GTP, the factor formed a ternary complex which would bind the 40S ribosomal subunits. 4. 4. Treatment of the factor with N-ethylmaleimide resulted in a loss of activity. 5. 5. Two sulfhydryl groups per eIF-2 molecule were essential for activity.
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