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Rapid Kinetic Analysis of Protein Synthesis

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Biophysical approaches to translational control of gene expression

Part of the book series: Biophysics for the Life Sciences ((BIOPHYS,volume 1))

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Abstract

Transient kinetic techniques provide information about chemical reactions, compositional dynamics, and conformational rearrangements of complex macromolecular assemblages. The use of those techniques has proven to be crucial in deciphering mechanisms of translation. To date, essentially every step of protein synthesis has been probed by rapid kinetic techniques. Understanding the functional mechanisms required specific methods to determine the timing and order of events, identify crucial intermediates, and understand the chemical and dynamic nature of the reactions involved. Here we describe the basics of rapid kinetic analysis and present two case studies that demonstrate how quench-flow and stopped-flow studies contribute to solving complex mechanisms. Analysis of chemical reactions at the active site of the ribosome revealed catalytic mechanisms of peptide bond formation and peptide release, while the analysis of conformational dynamics using fluorescence reporters provided insights into the function of elongation factor G in tRNA-mRNA translocation and ribosome recycling.

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Acknowledgments

The work in our labs was supported by the Deutsche Forschungsgemeinschaft and the Max Planck Society.

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Authors and Affiliations

  1. Max Planck Institute for Biophysical Chemistry, 37077, Goettingen, Germany

    Marina V. Rodnina & Wolfgang Wintermeyer

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  1. Marina V. Rodnina
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  2. Wolfgang Wintermeyer
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Correspondence to Marina V. Rodnina .

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Editors and Affiliations

  1. College Park, Cell Biology and Molecular Genetics, University of Maryland, Rm. 2135 Microbiology Building, College Park, 20742-4451, Maryland, USA

    Jonathan D. Dinman

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Rodnina, M.V., Wintermeyer, W. (2012). Rapid Kinetic Analysis of Protein Synthesis. In: Dinman, J. (eds) Biophysical approaches to translational control of gene expression. Biophysics for the Life Sciences, vol 1. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-3991-2_7

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