2v8x: Difference between revisions
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==Overview== | ==Overview== | ||
Structural complexes of the eukaryotic translation initiation factor 4E | Structural complexes of the eukaryotic translation initiation factor 4E (eIF4E) with a series of N(7)-alkylated guanosine derivative mRNA cap analogue structures have been characterised. Mass spectrometry was used to determine apparent gas-phase equilibrium dissociation constants (K(d)) values of 0.15 microM, 13.6 microM, and 55.7 microM for eIF4E with 7-methyl-GTP (m(7)GTP), GTP, and GMP, respectively. For tight and specific binding to the eIF4E mononucleotide binding site, there seems to be a clear requirement for guanosine derivatives to possess both the delocalised positive charge of the N(7)-methylated guanine system and at least one phosphate group. We show that the N(7)-benzylated monophosphates 7-benzyl-GMP (Bn(7)GMP) and 7-(p-fluorobenzyl)-GMP (FBn(7)GMP) bind eIF4E substantially more tightly than non-N(7)-alkylated guanosine derivatives (K(d) values of 7.0 microM and 2.0 microM, respectively). The eIF4E complex crystal structures with Bn(7)GMP and FBn(7)GMP show that additional favourable contacts of the benzyl groups with eIF4E contribute binding energy that compensates for loss of the beta and gamma-phosphates. The N(7)-benzyl groups pack into a hydrophobic pocket behind the two tryptophan side-chains that are involved in the cation-pi stacking interaction between the cap and the eIF4E mononucleotide binding site. This pocket is formed by an induced fit in which one of the tryptophan residues involved in cap binding flips through 180 degrees relative to structures with N(7)-methylated cap derivatives. This and other observations made here will be useful in the design of new families of eIF4E inhibitors, which may have potential therapeutic applications in cancer. | ||
==About this Structure== | ==About this Structure== | ||
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==Reference== | ==Reference== | ||
Crystallographic and | Crystallographic and mass spectrometric characterisation of eIF4E with N7-alkylated cap derivatives., Brown CJ, McNae I, Fischer PM, Walkinshaw MD, J Mol Biol. 2007 Sep 7;372(1):7-15. Epub 2007 Jun 15. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17631896 17631896] | ||
[[Category: Homo sapiens]] | [[Category: Homo sapiens]] | ||
[[Category: Protein complex]] | [[Category: Protein complex]] | ||
[[Category: Brown, C | [[Category: Brown, C J.]] | ||
[[Category: Fischer, P | [[Category: Fischer, P M.]] | ||
[[Category: Mcnae, I.]] | [[Category: Mcnae, I.]] | ||
[[Category: Walkinshaw, M | [[Category: Walkinshaw, M D.]] | ||
[[Category: MGQ]] | [[Category: MGQ]] | ||
[[Category: 4e-bp1]] | [[Category: 4e-bp1]] | ||
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[[Category: translation regulation]] | [[Category: translation regulation]] | ||
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Revision as of 19:54, 21 February 2008
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CRYSTALLOGRAPHIC AND MASS SPECTROMETRIC CHARACTERISATION OF EIF4E WITH N7-CAP DERIVATIVES
OverviewOverview
Structural complexes of the eukaryotic translation initiation factor 4E (eIF4E) with a series of N(7)-alkylated guanosine derivative mRNA cap analogue structures have been characterised. Mass spectrometry was used to determine apparent gas-phase equilibrium dissociation constants (K(d)) values of 0.15 microM, 13.6 microM, and 55.7 microM for eIF4E with 7-methyl-GTP (m(7)GTP), GTP, and GMP, respectively. For tight and specific binding to the eIF4E mononucleotide binding site, there seems to be a clear requirement for guanosine derivatives to possess both the delocalised positive charge of the N(7)-methylated guanine system and at least one phosphate group. We show that the N(7)-benzylated monophosphates 7-benzyl-GMP (Bn(7)GMP) and 7-(p-fluorobenzyl)-GMP (FBn(7)GMP) bind eIF4E substantially more tightly than non-N(7)-alkylated guanosine derivatives (K(d) values of 7.0 microM and 2.0 microM, respectively). The eIF4E complex crystal structures with Bn(7)GMP and FBn(7)GMP show that additional favourable contacts of the benzyl groups with eIF4E contribute binding energy that compensates for loss of the beta and gamma-phosphates. The N(7)-benzyl groups pack into a hydrophobic pocket behind the two tryptophan side-chains that are involved in the cation-pi stacking interaction between the cap and the eIF4E mononucleotide binding site. This pocket is formed by an induced fit in which one of the tryptophan residues involved in cap binding flips through 180 degrees relative to structures with N(7)-methylated cap derivatives. This and other observations made here will be useful in the design of new families of eIF4E inhibitors, which may have potential therapeutic applications in cancer.
About this StructureAbout this Structure
2V8X is a Protein complex structure of sequences from Homo sapiens with as ligand. Known structural/functional Site: . Full crystallographic information is available from OCA.
ReferenceReference
Crystallographic and mass spectrometric characterisation of eIF4E with N7-alkylated cap derivatives., Brown CJ, McNae I, Fischer PM, Walkinshaw MD, J Mol Biol. 2007 Sep 7;372(1):7-15. Epub 2007 Jun 15. PMID:17631896
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