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Updated: Jul 11, 2026

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
Published on: September 15, 2020
Protection-based assays to measure aminoacyl-tRNA binding to translation initiation factors
Yves Mechulam1, Laurent Guillon, Laure Yatime
1Laboratoire de Biochimie, CNRS Ecole Polytechnique, Palaiseau Cedex, France.
Abstract:
To decipher the mechanisms of translation initiation, the stability of the complexes between tRNA and initiation factors has to be evaluated in a routine manner. A convenient method to measure the parameters of binding of an aminoacyl-tRNA to an initiation factor results from the property that, when specifically complexed to a protein, the aminoacyl-tRNA often resists spontaneous deacylation. This chapter describes the preparation of suitable aminoacyl-tRNA ligands and their use in evaluating the stability of their complexes with various initiation factors, such as e/aIF2 and e/aIF5B. The advantages and the limitations of the method are discussed.
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