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

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  • Yves Mechulam
  • ,
  • Laurent Guillon
  • ,
  • Laure Yatime, Danmark
  • Sylvain Blanquet
  • ,
  • Emmanuelle Schmitt
  • Interdisciplinary Nanoscience Center
  • Molekylærbiologisk Institut
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.
OriginalsprogEngelsk
BogserieMethods in Enzymology
Vol/bind430
Sider (fra-til)265-81
Antal sider17
ISSN0076-6879
DOI
StatusUdgivet - 1 jan. 2007

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