Association of a cyclic AMP-dependent protein kinase with a purified translational inhibitor isolated from hemin-deficient rabbit reticulocyte lysates.
In the absence of added hemin, protein synthesis in rabbit reticulocyte lysates proceeds at maximal linear rates for several minutes and then ceases abruptly. Inhibition involves the action of a translational inhibitor whose formation is regulated by hemin. Addition of the isolated inhibitor to hemin-supplemented lysates produces an inhibition of protein chain initiation similar to that observed in heme-deficiency. The inhibitor has been purified over 300-fold and contains a protein kinase activity that copurifies with the inhibitory function. With calf thymus histone II as the phosphate receptor, the inhibitor-associated protein kinase requires ATP as the phosphorylating agent. Cycle AMP stimulates kinase activity 5- to 8-fold; the concentration of cycle AMP required for halfmaximal activity is 4 X 10-8 M. Preincubation of the inhibitor in the presence of cyclic AMP significantly reduces cyclic AMP-dependent phosphorylation and inhibitory activity. The corresponding protein kinase activity from hemin-supplemented lysates displays reduced cyclic AMP-dependency and little or no inhibitory activity. These findings suggest that the protein kinase activity associated with the purified translational inhibitor is involved in the mechanism of inhibition of initiation observed in hemedeficient reticulocyte lysates.
D000818:Animals; D001665:Binding Sites; D001798:Blood Proteins / Q000494:pharmacology*; D000242:Cyclic AMP / Q000494:pharmacology*; D006152:Cyclic GMP / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D006418:Heme / Q000031:analogs & derivatives*; D006427:Hemin / Q000494:pharmacology; D007527:Isoenzymes / Q000097:blood; D007700:Kinetics; D046911:Macromolecular Substances; D011485:Protein Binding; D014176:Protein Biosynthesis / Q000187:drug effects*; D011494:Protein Kinases / Q000097:blood*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism*
D001798:Blood Proteins; D007527:Isoenzymes; D046911:Macromolecular Substances; D006418:Heme; D006427:Hemin; D000242:Cyclic AMP; D011494:Protein Kinases; D006152:Cyclic GMP