Abstract
Eukaryotic initiation factor (eIF) 4F plays a key role in recruiting 40S ribosomes and associated factors to mRNA templates during translation initiation. The function of this heterotrimeric complex is to deliver an RNA helicase to the 5 cap proximal region of mRNAs in preparation for ribosome binding. To study the interaction between subunits of this complex, as well as identify small molecules that could interfere with their association, we developed a time resolved fluorescence assay that allows monitoring of interactions between two subunits of eIF4F. We have performed a small molecule chemical screen of > 73,000 compounds using this assay.
Keywords: Time-resolved fluorescent resonance energy transfer, FRET signal, Isothermal Titration Calorimetry, Recombinant Proteins, Eukaryotic initiation factor 4E binding protein
Combinatorial Chemistry & High Throughput Screening
Title: Homogenous Time Resolved Fluorescence Assay to Identify Modulators of Cap-Dependent Translation Initiation
Volume: 10 Issue: 3
Author(s): Regina Cencic, Yifei Yan and Jerry Pelletier
Affiliation:
Keywords: Time-resolved fluorescent resonance energy transfer, FRET signal, Isothermal Titration Calorimetry, Recombinant Proteins, Eukaryotic initiation factor 4E binding protein
Abstract: Eukaryotic initiation factor (eIF) 4F plays a key role in recruiting 40S ribosomes and associated factors to mRNA templates during translation initiation. The function of this heterotrimeric complex is to deliver an RNA helicase to the 5 cap proximal region of mRNAs in preparation for ribosome binding. To study the interaction between subunits of this complex, as well as identify small molecules that could interfere with their association, we developed a time resolved fluorescence assay that allows monitoring of interactions between two subunits of eIF4F. We have performed a small molecule chemical screen of > 73,000 compounds using this assay.
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Cite this article as:
Cencic Regina, Yan Yifei and Pelletier Jerry, Homogenous Time Resolved Fluorescence Assay to Identify Modulators of Cap-Dependent Translation Initiation, Combinatorial Chemistry & High Throughput Screening 2007; 10 (3) . https://dx.doi.org/10.2174/138620707780126688
DOI https://dx.doi.org/10.2174/138620707780126688 |
Print ISSN 1386-2073 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5402 |
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