Protection-based assays to measure aminoacyl-tRNA binding to translation initiation factors.

Yves Mechulam 1 Laurent Guillon 1 Laure Yatime 1 Sylvain Blanquet 1 Emmanuelle Schmitt 1, *
* Corresponding author
1 BIOC - Laboratoire de Biochimie de l'Ecole polytechnique
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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Submitted on : Tuesday, July 13, 2010 - 10:31:54 AM
Last modification on : Thursday, April 11, 2019 - 1:10:13 PM

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Yves Mechulam, Laurent Guillon, Laure Yatime, Sylvain Blanquet, Emmanuelle Schmitt. Protection-based assays to measure aminoacyl-tRNA binding to translation initiation factors.. Methods in Enzymology, Elsevier, 2007, 430, pp.265-81. ⟨10.1016/S0076-6879(07)30011-6⟩. ⟨hal-00502061⟩

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