University of Cambridge > Talks.cam > MRC LMB Seminar list > mRNA-tRNA translocation intermediates by cryo-EM: a look at the free-energy landscape from a single sample.

mRNA-tRNA translocation intermediates by cryo-EM: a look at the free-energy landscape from a single sample.

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After peptide bond formation (i.e., in the pre-translocational phase), the translating ribosome exists in multiple states that are in equilibrium. The states are characterized by different degrees of the ratchet-like intersubunit movement and different positions of the tRNAs, from classical to fully hybrid. Interconversion between these states is effected by Brownian motion, and the productive “end state” is fixed by the intervention of EF-G. Using cryo-EM of a single sample of the pre-translocational ribosome complex in the absence of EF-G, we were able to reconstruct the ribosome-tRNA configuration in several states. The varying ribosome domain and tRNA positions are quantified by tensor analysis and compared with those in the literature. A one-dimensional free-energy profile can be deduced from the relative occupancies of the states.

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