A fluorescence spectroscopic study on the binding of mRNA 5'-cap-analogs to human translation initiation factor eIF4E: a critical evaluation of the sources of error

J Photochem Photobiol B. 1998 May 15;43(2):158-63. doi: 10.1016/S1011-1344(98)00100-6.

Abstract

Equilibrium constants for the association of human protein translation initiation factor eIF4E with two mRNA 5'-cap analogs, namely 7-methylguanosine 5'-triphosphate and P1-(7-methylguanosine-5') P3-(guanosine-5') triphosphate, and with guanosine 5'-monophosphate have been redetermined by the fluorescence quenching method taking the inner filter effect of the cap-analog into account. It has been shown that neglecting the latter correction may lead to either underestimation or overestimation of the association constant obtained by applying the Eadie-Hofstee plot: the reasonably firm binding of 7-methylated cap-analogs becomes underestimated, while the weak binding of non-methylated nucleotides becomes overestimated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Erythrocytes / metabolism
  • Escherichia coli
  • Eukaryotic Initiation Factor-4E
  • Humans
  • Kinetics
  • Methylation
  • Peptide Initiation Factors / metabolism*
  • RNA Caps / chemistry
  • RNA Caps / metabolism*
  • RNA, Messenger / chemistry
  • RNA, Messenger / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Reproducibility of Results
  • Spectrometry, Fluorescence / methods
  • Structure-Activity Relationship

Substances

  • Eukaryotic Initiation Factor-4E
  • Peptide Initiation Factors
  • RNA Caps
  • RNA, Messenger
  • Recombinant Proteins