Fig. 5: The ScIleRS·tRNAIle(CAU)·l-Ile complex is stalled in a non-reactive state due to the lack of key N34 interactions. | Nature Communications

Fig. 5: The ScIleRS·tRNAIle(CAU)·l-Ile complex is stalled in a non-reactive state due to the lack of key N34 interactions.

From: The mechanism of discriminative aminoacylation by isoleucyl-tRNA synthetase based on wobble nucleotide recognition

Fig. 5

a The aminoacylation activities of ScIleRS and SaMetRS against in vitro transcribed tRNAIle(GAU), tRNAIle(CAU) and tRNAIle(CAU) C34G variant. Data are presented as means ± SD (n = 3 independent experiments). b EMSA revealed that in vitro transcribed tRNAIle(CAU) can still form a complex with ScIleRS, although it is slightly weaker than in vitro transcribed tRNAIle(GAU). Similar results were observed in two independent experiments. c The overall structure of the ScIleRS·tRNAIle(CAU)·l-Ile complex. The anticodon loop of tRNAIle(CAU) is dynamic because of the lack of interactions with ScIleRS, and the acceptor stem of tRNAIle(CAU) binds to the back of the ED. d Binding of the acceptor stem of tRNAIle(CAU) to the back of ScIleRS ED. The 3’ CCA end was invisible in the electronic density map. e Structural comparison between the reactive and non-reactive ScIleRS·tRNA complexes revealed an approximately 25° rotation between tRNAIle(GAU) and tRNAIle(CAU). f The C-terminal domains of tRNAIle(CAU)-bound ScIleRS adopt a conformation generally similar to that of ScIleRS in the tRNA-free state except for the distal C-ter C ___domain, but not to that of tRNAIle(GAU)-bound ScIleRS.

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