Effect of antibody to the hemin-controlled translational repressor in rabbit reticulocyte lysate.
We have examined the effect of the purified IgG from the serum of guinea pigs immunized with a highly purified preparation of rabbit reticulocyte, hemin-controlled translational repressor (HCR) on protein synthesis in the reticulocyte lysate. We have found that the anti-HCR (but not non-immune) IgG completely prevents or reverses the suppression of protein synthesis that occurs in hemin-deficient lysate, providing a direct and definitive demonstration that the inhibitory effect of hemin-deficiency is mediated solely by the activation of HCR. The anti-HCR IgG also prevents or reverses the phosphorylation of eIF-2 alpha and the reduced binding of Met-tRNAf to 40 S ribosomal subunits that accompanies the inhibition of protein synthesis in hemin-deficient lysate. In contrast, the anti-HCR IgG has no effect on the inhibition produced by low levels of double-stranded RNA (that is due to the activation of a separate protein kinase), but it does partly reverse inhibition due to oxidized glutathione, ethanol, and phosphatidylserine, indicating that the effect of these components is mediated, at least in part, by the activation of HCR. Finally, we have confirmed our earlier observation that an excess of proHCR, the inactive precursor of HCR, has little effect on the neutralization of HCR by limiting anti-HCR IgG, suggesting that the antigenic determinants on HCR are not exposed on ProHCR.
D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D003603:Cytotoxins; D006427:Hemin / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D011494:Protein Kinases / Q000096:biosynthesis / Q000276:immunology*; D011817:Rabbits; D012156:Reticulocytes / Q000187:drug effects*; D013696:Temperature; D019892:eIF-2 Kinase
D000906:Antibodies; D003603:Cytotoxins; D007074:Immunoglobulin G; D006427:Hemin; D011494:Protein Kinases; D019892:eIF-2 Kinase