The activity of eukaryotic initiation factor eIF-2 in ternary complex formation with GTP and Met-tRNA.
Ternary complex formation between [3H]Met-tRNAf, [14C]H3-eIF-2, and GTP was measured on nitrocellulose filters. It is shown that [3H]Met-tRNAf and [14C]H3-eIF-2 are present on the filter in equimolar amounts when ATP, creatine phosphate, and creatine kinase are included in the reaction mixture. Under these conditions the factor is 100% active. With small amounts of factor significant losses occur due to adsorption to the wall of the reaction vessels, resulting in seemingly low activities of eIF-2. These losses can be prevented by the presence of "stimulatory" proteins, which enhance the recovery of both [3H]Met-tRNAf and [14C]H3-eIF-2 on the filter but do not alter their ratio.
D000818:Animals; D006160:Guanosine Triphosphate / Q000378:metabolism*; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D046911:Macromolecular Substances; D008715:Methionine; D010442:Peptide Chain Initiation, Translational; D010448:Peptide Initiation Factors / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism*; D051381:Rats; D012156:Reticulocytes / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
D046911:Macromolecular Substances; D010448:Peptide Initiation Factors; D006160:Guanosine Triphosphate; D012343:RNA, Transfer; D008715:Methionine