Binding and release of eukaryotic initiation factor eIF-2 and GTP during protein synthesis initiation.
The eukaryotic initiation factor eIF-2 forms a ternary complex with Met-tRNAf and GTP. This complex binds to the 40S ribosomal subunit in the absence of mRNA and mRNA binding factors. Highly purified eIF-2 from rabbit reticulocytes was labeled with 125I by using the Bolton-Hunter reagent or with [gamma-32P]ATP by using the heme-regulated translational inhibitor protein kinase. The labeled eIF-2 was bound, together with equimolar amounts of Met-tRNAf and GTP, to the 40S subunit. In the presence of mRNA, mRNA binding factors, and 60S ribosomal subunits (complete initiation assay), eIF-2 was released from the 40S initiation complex in the subunit joining reaction. GTP also was released in this step and probably was hydrolyzed in the reaction that is dependent upon eIF-5 and the 60S subunit. The function of phosphorylated eIF-2 in initiation of protein synthesis is discussed.
D000818:Animals; D006160:Guanosine Triphosphate / Q000378:metabolism*; D010442:Peptide Chain Initiation, Translational; D010448:Peptide Initiation Factors; D010710:Phosphates / Q000378:metabolism; D011485:Protein Binding; D012343:RNA, Transfer / Q000378:metabolism; D011817:Rabbits; D012270:Ribosomes / Q000378:metabolism*
D010448:Peptide Initiation Factors; D010710:Phosphates; D006160:Guanosine Triphosphate; D012343:RNA, Transfer
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