Abstract <p>The review discusses the functional role of the ribosomal E-site in the context of recent structural data. Traditionally, the E-site has been considered to serve only as a binding site for deacylated tRNA (E-tRNA) prior to its dissociation from the protein synthesis complex. Here, we examine specific contacts formed between E-tRNA and rRNA of the large ribosomal subunit in different organisms, as well as the sequence of their formation and disruption. The mechanism of translation suppression by inhibitors that bind to the ribosomal E-site is discussed. Based on current evidence regarding the location of aminoacyl-tRNA synthetases (ARSs) in the immediate vicinity of the ribosome, we propose a hypothesis that one of the primary functions of the ribosomal E-site is to prepare tRNA (through its modulation) for the formation of a specific complex with ARS, in the content of which it is released from the ribosome.</p>