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bioRxiv · 10.1101/2022.10.03.510626

Entropic control of the free energy landscape of an archetypal biomolecular machine

Ray, K. K.·Kinz-Thompson, C. D.·Fei, J.·Wang, B.·Lin, Q.·Gonzalez, R. L.
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Abstract

Biomolecular machines are complex macromolecular assemblies that utilize thermal and chemical energy to perform essential, multistep, cellular processes. Despite possessing different architectures and functions, an essential feature of the mechanisms-of-action of all such machines is that they require dynamic rearrangements of structural components. Surprisingly, biomolecular machines generally possess only a limited set of such motions, suggesting that these dynamics must be repurposed to drive different mechanistic steps. Although ligands that interact with these machines are known to drive such repurposing, the physical and structural mechanisms through which ligands achieve this remain unknown. Using temperature-dependent, single-molecule measurements analyzed with a time-resolution-enhancing algorithm, here we dissect the free energy landscape of an archetypal biomolecular machine, the bacterial ribosome, to reveal how its dynamics are repurposed to drive distinct steps during ribosome-catalyzed protein synthesis. Specifically, we show that the free energy landscape of the ribosome encompasses a network of allosterically coupled structural elements that coordinates the motions of these elements. Moreover, we reveal that ribosomal ligands which participate in disparate steps of the protein synthesis pathway repurpose this network by differentially modulating the structural flexibility of the ribosomal complex (i.e., the entropic component of the free energy landscape). We propose that such ligand-dependent entropic control of free energy landscapes has evolved as a general strategy through which ligands may regulate the functions of all biomolecular machines. Such entropic control is therefore an important driver in the evolution of naturally occurring biomolecular machines and a critical consideration for the design of synthetic molecular machines.

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Ray, K. K., Kinz-Thompson, C. D., Fei, J., Wang, B., Lin, Q., Gonzalez, R. L.. 2022-10-05. Entropic control of the free energy landscape of an archetypal biomolecular machine. https://doi.org/10.1101/2022.10.03.510626

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