Microtubule-Associated Proteins and Tubulin Interaction by Isothermal Titration Calorimetry

Philipp O Tsvetkov (1) , Pascale Barbier (1) , Gilles Breuzard (1) , Vincent Peyrot (1) , François Devred (1)

1 CRO2 - Centre de Recherches en Oncologie biologique et Oncopharmacologie

Microtubules play an important role in a number of vital cell processes such as cell division, intracellular transport, and cell architecture. The highly dynamic structure of microtubules is tightly regulated by a number of stabilizing and destabilizing microtubule-associated proteins (MAPs), such as tau and stathmin. Because of their importance, tubulin–MAPs interactions have been extensively studied using various methods that provide researchers with complementary but sometimes contradictory thermodynamic data. Isothermal titration calorimetry (ITC) is the only direct thermodynamic method that enables a full thermodynamic characterization (stoichiometry, enthalpy, entropy of binding, and association constant) of the interaction after a single titration experiment. This method has been recently applied to study tubulin–MAPs interactions in order to bring new insights into molecular mechanisms of tubulin regulation. In this chapter, we review the technical specificity of this method and then focus on the use of ITC in the investigation of tubulin– MAPs binding. We describe technical issues which could arise during planning and carrying out the ITC experiments, in particular with fragile proteins such as tubulin. Using examples of stathmin and tau, we demonstrate how ITC can be used to gain major insights into tubulin–MAP interaction.

Mots clés: ITC; microtubule; tubulin; MAP; microtubule associated protein; tau; isothermal titration calorimetry; stathmin

Domaines: Cancer; Neurobiologie; Biophysique

Philipp O Tsvetkov, Pascale Barbier, Gilles Breuzard, Vincent Peyrot, François Devred. Microtubule-Associated Proteins and Tubulin Interaction by Isothermal Titration Calorimetry. Methods in Cell Biology, 115, pp.283 - 302, 2013, Microtubules, In Vitro 978-0-12-407757-7. ⟨10.1016/B978-0-12-407757-7.00018-9⟩. ⟨hal-01478645⟩