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)

1 BIOC - Laboratoire de Biochimie de l'Ecole polytechnique

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.

Domains: Life Sciences [q-bio]; Biochemistry, Molecular Biology

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, 2007, 430, pp.265-81. ⟨10.1016/S0076-6879(07)30011-6⟩. ⟨hal-00502061⟩