pH-dependent and ligand induced conformational changes of eucaryotic protein synthesis initiation factor eIF-(iso)4F: a circular dichroism study

Biochim Biophys Acta. 1996 Oct 17;1297(2):207-13. doi: 10.1016/s0167-4838(96)00119-7.

Abstract

The structural features of wheat germ protein synthesis initiation factor eIF-(iso)4F, which has a cap binding protein as one of its two subunits, are unknown. In this study, circular dichroism (CD) spectra and secondary structure prediction were obtained for eIF-(iso)4F and its two subunits, p28 and p86. The alpha-helix content of eIF-(iso)4F changed from 42% at pH 6.3 to 15% at pH 7.6, the optimum pH for cap binding. The beta-sheet content increased from 14% (pH 6.3) to 38% at pH 7.6. The CD spectra of the two subunits, p28 and p86 were also measured and analyzed. The separated subunits both had a higher alpha-helix content at pH 7.6 than the native protein, giving values of 60% and 34% alpha-helix for p28 and p86, respectively. Binding of the dinucleotide cap analog to p28 reduced the alpha-helix content to approximately 8% with an increase in the beta sheet content from 10% to 37%. The conformational changes in eIF-(iso)4F upon binding with mRNA are dependent on cap or oligonucleotide structure. A conformation consisting of approximately the same alpha-helix and beta-sheet content can be induced by ligands even at non-optimal pH values. This large conformational transition suggests eIF-(iso)4F binds nucleic acids by interaction of a beta-sheet motif and that this conformational transition may have a regulatory role.

Publication types

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

MeSH terms

  • Circular Dichroism
  • Eukaryotic Initiation Factor-1 / chemistry*
  • Eukaryotic Initiation Factor-1 / metabolism*
  • Hydrogen-Ion Concentration
  • Ligands
  • Nucleic Acid Conformation
  • Peptide Initiation Factors / chemistry*
  • Peptide Initiation Factors / metabolism*
  • Protein Conformation*
  • Protein Structure, Secondary
  • RNA Cap-Binding Proteins
  • RNA-Binding Proteins / chemistry
  • Triticum / chemistry*

Substances

  • Eukaryotic Initiation Factor-1
  • Ligands
  • Peptide Initiation Factors
  • RNA Cap-Binding Proteins
  • RNA-Binding Proteins
  • eIF-4