Assay for Identification of Therapeutics Targeting Ternary Complex Formation in Protein Synthesis

摘要

The present invention provides a novel assay that allows high-throughput screening of chemical compounds for the inhibition of binding between EF-Tu and tRNA.

主权项

1. A method of identifying whether a test compound possesses antimicrobial activity, said method comprising:
a) mixing an elongation factor Tu (EF-Tu) with GTP and an aminoacylated transfer RNA (aa-tRNA) in the presence of said test compound, wherein said EF-Tu is operably linked to a first energy transfer pair member, wherein said aa-tRNA is operably linked to a second energy transfer pair member, and wherein said first energy transfer pair member and said second energy transfer pair member form a fluorescence resonance energy transfer pair; and b) detecting the fluorescence emitted, wherein the level of fluorescence in the presence of the test compound is compared to the level of fluorescence in the absence of the test compound or the level of fluorescence in the presence of a control compound, wherein a difference in fluorescence indicates that the test compound possesses antimicrobial activity.