Structure and function of peptide initiation factor 2: differential loss of activities during proteolysis and generation of a terminal fragment containing the phosphorylation sites of the alpha subunit.
A procedure is described by which the 38,000-dalton alpha subunit of native eukaryotic peptide initiation factor 2 (eIF-2) can be cleaved by trypsin to yield a 34,000-dalton fragment and a peptide of about 4,000 daltons after elimination of the beta subunit. Under nondenaturing conditions the 4,000-dalton peptide remains bound to the modified eIF-2 and still can be phosphorylated by the heme-controlled eIF-2 alpha kinase from reticulocytes. All of the phosphorylation sites for this protein kinase are located on the 4,000-dalton peptide. The ability of eIF-2 to form a ternary complex with GTP and Met-tRNAf and the ability to promote binding of Met-tRNAf to 40S ribosomal subunits are lost differentially during the proteolysis. Loss of te latter activity occurs rapidly and appears to be correlated with loss of the beta subunit. Loss of activity for ternary complex formation is correlated with the appearance of the 4,000-dalton peptide.
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002474:Cell-Free System; D015852:Eukaryotic Initiation Factor-2; D006160:Guanosine Triphosphate / Q000378:metabolism; D006868:Hydrolysis; D046911:Macromolecular Substances; D010442:Peptide Chain Initiation, Translational; D010446:Peptide Fragments / Q000378:metabolism; D010448:Peptide Initiation Factors / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D011817:Rabbits; D013329:Structure-Activity Relationship; D019892:eIF-2 Kinase
D015852:Eukaryotic Initiation Factor-2; D046911:Macromolecular Substances; D010446:Peptide Fragments; D010448:Peptide Initiation Factors; D011506:Proteins; D012346:RNA, Transfer, Amino Acyl; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D011494:Protein Kinases; D019892:eIF-2 Kinase