Protein synthesis in brine shrimp embryos and rabbit reticulocytes. The effect of Mg2+ on binary (eukaryotic initiation factor 2 X GDP) and ternary (eukaryotic initiation factor 2 X GTP X met-tRNAf) complex formation

J Biol Chem. 1983 Mar 25;258(6):3438-41.

Abstract

We have prepared eukaryotic initiation factor 2 (eIF-2) from rabbit reticulocytes and Artemia embryos and studied the effect of Mg2+ on binary (eIF-2 X GDP) and ternary (eIF-2 X GTP X Met-tRNAf) complex formation. Under conditions where Mg2+ inhibits Met-tRNAf binding to reticulocyte eIF-2, ternary complex formation with Artemia eIF-2 is not inhibited. Similarly, the formation of eIF-2 X GDP with Artemia eIF-2 is stimulated by Mg2+, whereas the corresponding reticulocyte binary complex is strongly inhibited. In the presence of 1 mM Mg2+, the isolated Artemia eIF-2 X GDP complex is stable in the absence of any added nucleotide, but readily exchanges bound GDP for free GTP. However, the reticulocyte eIF-2 X GDP complex is significantly more stable in the presence of GTP, and nucleotide exchange is dependent upon the addition of a factor isolated from either the postribosomal supernatant or the high salt wash of rabbit reticulocyte ribosomes. This factor also stimulates Met-tRNAf binding to both Artemia and reticulocyte eIF-2.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Artemia / metabolism*
  • Embryo, Nonmammalian / metabolism*
  • Eukaryotic Initiation Factor-2
  • Guanosine Diphosphate / metabolism
  • Guanosine Triphosphate / metabolism
  • Kinetics
  • Magnesium / pharmacology
  • Peptide Chain Initiation, Translational* / drug effects
  • Peptide Initiation Factors / metabolism*
  • Proteins / metabolism*
  • RNA, Transfer, Amino Acyl / metabolism
  • RNA, Transfer, Met*
  • Rabbits
  • Reticulocytes / metabolism*

Substances

  • Eukaryotic Initiation Factor-2
  • Peptide Initiation Factors
  • Proteins
  • RNA, Transfer, Amino Acyl
  • RNA, Transfer, Met
  • tRNA, formylmethionine-
  • Guanosine Diphosphate
  • Guanosine Triphosphate
  • Magnesium