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Thermodynamics of Molecular Machines Using Incremental ITC

  • Benoît Meyer
  • Cyrielle da Veiga
  • Philippe Dumas
  • Eric EnnifarEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1964)

Abstract

Molecular biomachines, such as DNA and RNA polymerases or the ribosome, are fascinating biological assemblies able to swiftly perform repeated and highly regulated tasks, with a remarkable accuracy. Significant advances in structural studies during the past 20 years provided a wealth of information regarding their architecture and considerably contributed to a better understanding of their mechanism of action. However, the three-dimensional structure of a biological nanomachine alone does not provide access to its detailed mechanism of action, even when obtained at atomic resolution. When combined with other biophysical approaches, thermodynamic data, together with kinetic data, are essential for a complete description of any binding interaction, revealing forces driving complex formation and providing insights into mechanisms of action. We have developed an incremental ITC approach that is well-suitable for analysis of biomolecular machines. This strategy allows a dissection of molecular biomachine reactions through successive additions in the ITC cell of consecutive substrates.

Key words

HIV reverse transcriptase Incremental ITC Thermodynamics Molecular biomachines Mechanism of action 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Benoît Meyer
    • 1
  • Cyrielle da Veiga
    • 1
  • Philippe Dumas
    • 1
  • Eric Ennifar
    • 1
    Email author
  1. 1.Institut de Biologie Moléculaire et CellulaireUniversité de Strasbourg, CNRSStrasbourgFrance

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