Abstract
Microtubules (MTs) play an important role in many cellular processes and are dynamic structures regulated by an important network of microtubules-associated proteins, MAPs, such as Tau. Tau has been discovered as an essential factor for MTs formation in vitro, and its region implicated in binding to MTs has been identified. By contrast, the affinity, the stoichiometry, and the topology of Tau–MTs interaction remain controversial. Indeed, depending on the experiment conditions a wide range of values have been obtained. In this chapter, we focus on three biophysical methods, turbidimetry, cosedimentation assay, and Förster Resonance Energy Transfer to study Tau–tubulin interaction both in vitro and in cell. We highlight precautions that must be taken in order to avoid pitfalls and we detail the nature of the conclusions that can be drawn from these methods about Tau–tubulin interaction.
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References
Mitchison T, Kirschner M (1984) Dynamic instability of microtubule growth. Nature 312:237–242
Bhat KM, Setaluri V (2007) Microtubule-associated proteins as targets in cancer chemotherapy. Clin Cancer Res 13:2849–2854
Weingarten MD, Lockwood AH, Hwo SY et al (1975) A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A 72:1858–1862
Tilney LG, Bryan J, Bush DJ et al (1973) Microtubules: evidence for 13 protofilaments. J Cell Biol 59:267–275
Chrétien D, Wade RH (1991) New data on the microtubule surface lattice. Biol Cell 71:161–174
Wade RH (2007) Microtubules: an overview. Methods Mol Med 137:1–16
Weisenberg RC, Timasheff SN (1970) Aggregation of microtubule subunit protein. Effects of divalent cations, colchicine and vinblastine. Biochemistry 9:4110–4116
Frigon RP, Timasheff SN (1975a) Magnesium-induced self-association of calf brain tubulin. I. Stoichiometry. Biochemistry 14:4559–4566
Frigon RP, Timasheff SN (1975b) Magnesium-induced self-association of calf brain tubulin. II. Thermodynamics. Biochemistry 14:4567–4573
Mandelkow EM, Mandelkow E, Milligan RA (1991) Microtubule dynamics and microtubule caps: a time-resolved cryo-electron microscopy study. J Cell Biol 114:977–991
Müller-Reichert T, Chrétien D, Severin F, Hyman AA (1998) Structural changes at microtubule ends accompanying GTP hydrolysis: information from a slowly hydrolyzable analogue of GTP, guanylyl (alpha, beta)methylenediphosphonate. Proc Natl Acad Sci U S A 95:3661–3666
Brouhard GJ, Rice LM (2014) The contribution of αβ-tubulin curvature to microtubule dynamics. J Cell Biol 207:323–334
Himmler A, Drechsel D, Kirschner MW, Martin DW Jr (1989) Tau consists of a set of proteins with repeated C-terminal microtubule-binding domains and variable N-terminal domains. Mol Cell Biol 9:1381–1388
Butner KA, Kirschner MW (1991) Tau protein binds to microtubules through a flexible array of disturbed weak sites. J Cell Biol 115:717–730
Goode BL, Feinstein SC (1994) Identification of a novel microtubule binding and assembly domain in the developmentally regulated inter-repeat region of tau. J Cell Biol 124:769–782
Gustke N, Trinczek B, Biernat J et al (1994) Domains of tau protein and interactions with microtubules. Biochemistry 33:9511–9522
Sillen A, Barbier P, Landrieu I et al (2007) NMR investigation of the interaction between the neuronal protein tau and the microtubules. Biochemistry 46:3055–3064
Devred F, Barbier P, Lafitte D et al (2010) Microtubule and MAPs: thermodynamics of complex formation by AUC, ITC, fluorescence, and NMR. Methods Cell Biol 95:449–480
Tsvetkov PO, Makarov AA, Malesinski S et al (2012) New insights into tau-microtubules interaction revealed by isothermal titration calorimetry. Biochimie 94:916–919
Tsvetkov PO, Barbier P, Breuzard G et al (2013) Microtubule-associated proteins and tubulin interaction by isothermal titration calorimetry. Methods Cell Biol 115:283–302
Goedert M, Wischik CM, Crowther RA et al (1988) Cloning and sequencing of the cDNA encoding a core protein of the paired helical filament of Alzheimer disease: identification as the microtubule-associated protein tau. Proc Natl Acad Sci U S A 85:4051–4055
Dompierre JP, Godin JD, Charrin BC et al (2007) Histone deacetylase 6 inhibition compensates for the transport deficit in Huntington’s disease by increasing tubulin acetylation. J Neurosci 27:3571–3583
Rasband W (1997–2007) ImageJ. US National Institutes of Health, Bethesda, MA, http://rsb.info.nih.gov/ij/plugins.html
Feige JN, Sage D, Wahli W et al (2005) PixFRET, an ImageJ plug-in for FRET calculation that can accommodate variations in spectral bleed-throughs. Microsc Res Tech 68:51–58
Oosawa F, Asakura S (1975) Thermodynamics of the polymerization of protein. Academic, London
Devred F, Douillard S, Briand C, Peyrot V (2002) First tau repeat domain to growing and taxol-stabilized microtubules, and serine 262 residue phosphorylation. FEBS Lett 523:247–251
Weis F, Moullintraffort L, Heichette C et al (2010) The 90-kDa heat shock protein Hsp90 protects tubulin against thermal denaturation. J Biol Chem 285:9525–9534
Devred F, Barbier P, Douillard S et al (2004) Tau induces ring and microtubule formation from alphabeta-tubulin dimers under nonassembly conditions. Biochemistry 43:10520–10531
Buey RM, Diaz JF, Andreu JM et al (2004) Interaction of epothilone analogues with the paclitaxel binding site: relationship between binding affinity, microtubule stabilization, and cytotoxicity. Chem Biol 11:225–236
Breuzard G, Hubert P, Nouar R et al (2013) Molecular mechanisms of Tau binding to microtubules and its role in microtubule dynamics in live cells. J Cell Sci 126:2810–2819
Wouters FS, Verveer PJ, Bastiaens PI (2001) Imaging biochemistry inside cells. Trends Cell Biol 11:203–211
Gordon GW, Berry G, Liang XH et al (1998) Quantitative fluorescence resonance energy transfer measurements using fluorescence microscopy. Biophys J 74:2702–2713
Xia Z, Liu Y (2001) Reliable and global measurement of fluorescence resonance energy transfer using fluorescence microscopes. Biophys J 81:2395–2402
Youvan DC, Silva CM, Bilina E et al (1997) Calibration of fluorescence resonance energy transfer in microscopy using genetically engineered GFP derivatives on nickel chelating beads. Biotechnology 3:1–18
Gonzales RC, Woods RE (2007) Digital image processing, 3rd edn. Pearson Education, Upper Saddle River, NJ
Pratt WK (2006) Digital image processing, 4th edn. Wiley-Interscience, New York, NY
Siegel RM, Chan FK, Zacharias DA et al (2000) Measurement of molecular interactions in living cells by fluorescence resonance energy transfer between variants of the green fluorescent protein. Sci STKE 38:1
Nouar R, Devred F, Breuzard G, Peyrot V (2013) FRET and FRAP imaging: approaches to characterise tau and stathmin interactions with microtubules in cells. Biol Cell 105:149–161
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De Bessa, T., Breuzard, G., Allegro, D., Devred, F., Peyrot, V., Barbier, P. (2017). Tau Interaction with Tubulin and Microtubules: From Purified Proteins to Cells. In: Smet-Nocca, C. (eds) Tau Protein. Methods in Molecular Biology, vol 1523. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6598-4_4
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DOI: https://doi.org/10.1007/978-1-4939-6598-4_4
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