title: Protection-based assays to measure aminoacyl-tRNA binding to translation initiation factors.
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.
authors.affiliation: Laboratoire de Biochimie, CNRS Ecole Polytechnique, Palaiseau Cedex, France.
authors.forename: Yves
authors.initials: Y
authors.lastname: Mechulam
authors.forename: Laurent
authors.initials: L
authors.lastname: Guillon
authors.forename: Laure
authors.initials: L
authors.lastname: Yatime
authors.forename: Sylvain
authors.initials: S
authors.lastname: Blanquet
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Schmitt
mesh_terms: D039642:Eukaryotic Initiation Factors / Q000378:metabolism*; D014176:Protein Biosynthesis; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D039642:Eukaryotic Initiation Factors; D012346:RNA, Transfer, Amino Acyl; D012358:RNA, Transfer, Met