Homogenous Time Resolved Fluorescence Assay to Identify Modulators of Cap-Dependent Translation Initiation

Author(s): Regina Cencic, Yifei Yan, Jerry Pelletier

Journal Name: Combinatorial Chemistry & High Throughput Screening
Accelerated Technologies for Biotechnology, Bioassays, Medicinal Chemistry and Natural Products Research

Volume 10 , Issue 3 , 2007

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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

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Article Details

VOLUME: 10
ISSUE: 3
Year: 2007
Page: [181 - 188]
Pages: 8
DOI: 10.2174/138620707780126688
Price: $65

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