Microtubule and MAPs: thermodynamics of complex formation by AUC, ITC, fluorescence, and NMR.
Microtubules are implicated in many essential cellular processes such as architecture, cell division, and intracellular traffic, due to their dynamic instability. This dynamicity is tightly regulated by microtubule-associated proteins (MAPs), such as tau and stathmin. Despite extensive studies motivated by their central role in physiological functions and pathological role in neurodegenerative diseases and cancer, the precise mechanisms of tau and stathmin binding to tubulin and their consequences on microtubule stability are still not fully understood. One of the most crucial points missing is a quantitative thermodynamic description of their interaction with tubulin/microtubules and of the tubulin complexes formed upon these interactions. In this chapter, we will focus on the use of analytical ultracentrifugation, isothermal titration calorimetry, and nuclear magnetic resonance-three powerful and complementary techniques in the field of MAP-tubulin/microtubule interactions, in addition to the spectrometric techniques and co-sedimentation approach. We will present the limits of these techniques to study this particular interaction and precautions that need to be taken during MAPs preparation. Understanding the molecular mechanisms that govern MAPs action on microtubular network will not only shed new light on the role of this crucial family of protein in the biology of the cell, but also hopefully open new paths to increase the therapeutic efficiency of microtubule-targeting drugs in cancers therapies and neurodegeneratives diseases prevention.
Devred
François
F
CRO2, U911 Inserm, Aix-Marseille Université, 27 Bd Jean Moulin, 13385 Marseille cedex 05, France.
Barbier
Pascale
P
Lafitte
Daniel
D
Landrieu
Isabelle
I
Lippens
Guy
G
Peyrot
Vincent
V
D000818:Animals; D002152:Calorimetry, Differential Scanning / Q000379:methods; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008869:Microtubule-Associated Proteins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods; D013816:Thermodynamics; D014461:Ultracentrifugation / Q000379:methods
D046911:Macromolecular Substances; D008869:Microtubule-Associated Proteins