Biology Reference
In-Depth Information
CHAPTER
18
Microtubule-Associated
Proteins and Tubulin
Interaction by Isothermal
Titration Calorimetry
P.O. Tsvetkov
*
, P. Barbier
{
, G. Breuzard
{
, V. Peyrot
{
and F. Devred
{
*
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Moscow, Russia
{
Aix-Marseille University, Inserm, CRO2 UMR_S 911, Faculte´ de Pharmacie,
13385 Marseille, France
CHAPTER OUTLINE
Introduction ............................................................................................................ 284
18.1 Isothermal Titration Calorimetry.......................................................................286
18.1.1 Principles .................................................................................. 286
18.1.2 Experimental Procedure .............................................................. 286
18.1.3 Binding Models .......................................................................... 287
18.1.4 Thermodynamic Profile of Binding (Meaning of Enthalpy
and Entropy Signs) ..................................................................... 289
18.1.5 Temperature Dependence of
.................................................. 289
18.2 Tubulin and MAPs SAMPLE PREPARATION ........................................................290
18.2.1 Equilibration .............................................................................. 290
18.2.2 Determination of Protein Concentrations....................................... 291
18.2.3 Temperature .............................................................................. 292
18.2.4 Buffer Conditions ....................................................................... 293
18.2.5 Reaction Volume and Duration..................................................... 293
18.3 Results: Tubulin/MAPs by ITC ..........................................................................294
18.3.1 Stathmin-Tubulin Interaction ...................................................... 295
18.3.2 Tau-Tubulin............................................................................... 297
Conclusion ............................................................................................................. 298
Acknowledgment..................................................................................................... 299
References ............................................................................................................. 299
D
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