Abstract
Stathmin is an important phosphorylation-controlled regulator of microtubule dynamics and plays a crucial role in cell division and cell proliferation. In its non-phosphorylated form, stathmin is the protein that interacts the most tightly with tubulin, in a 2:1 tubulin-stathmin (T2S) complex that does not participate in microtubule assembly. The importance of stathmin at different levels of phosphorylation in different steps of mitosis This article is a short overview of the different methods that have been or could be used to monitor the kinetic and thermodynamic parameters of tubulin-stathmin interaction and to evaluate the effects of phosphorylation. The author has tried to emphasize how hydrodynamic and spectroscopic methods measuring direct binding of stathmin to tubulin can be complemented by methods that make use of linked functions, measuring how the change in a functional property of tubulin upon binding stathmin provides information on binding parameters.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Wyman, J. (1984) Linkage graphs: a study in the thermodynamics of macromolecules. Q. Rev. Biophys. 17, 453–488.
Beretta, L., Dobransky, T., and Sobel, A. (1993) Multiple phosphorylation of stathmin. Identification of four sites phosphorylated in intact cells and in vitro by cyclic AMP-dependent protein kinase and p34cdc2. J. Biol. Chem. 268, 20,076–20,084.
Brattsand, G., Marklund, U., Nylander, K., Roos, G., and Gullberg, M. (1994) Cell-cycle-regulated phosphorylation of oncoprotein 18 on Ser16, Ser25 and Ser38. Eur. J. Biochem. 220, 359–368.
Larsson, N., Melander, H., Marklund, U., Osterman, O., and Gullberg, M. (1995) G2/M transition requires multisite phosphorylation of oncoprotein 18 by two distinct protein kinase systems. J. Biol. Chem. 270, 14,175–14,183.
Marklund, U., Brattsand, G., Osterman, O., Ohlsson, P. I., and Gullberg, M. (1993) Multiple signal transduction pathways induce phosphorylation of serines 16, 25, and 38 of oncoprotein 18 in T lymphocytes. J. Biol. Chem. 268, 25,671–25,680.
Marklund, U., Larsson, N., Gradin, H. M., Brattsand, G., and Gullberg, M. (1996) Oncoprotein 18 is a phosphorylation-responsive regulator of microtubule dynamics. EMBO J. 15, 5290–5298.
Andersen, S. S., Ashford, A. J., Tournebize, R., et al. (1997) Mitotic chromatin regulates phosphorylation of Stathmin/Op18. Nature 389, 640–643.
Laird, A. D. and Shalloway, D. (1997) Oncoprotein signalling and mitosis. Cell Signal. 9, 249–255.
Moreno, F. J. and Avila, J. (1999) Phosphorylation of stathmin modulates its function as a microtubule depolymerizing factor. Mol. Cell. Biochem. 183, 201–209.
Sobel, A. (1991) Stathmin: a relay phosphoprotein for multiple signal transduction? Trends Biochem. Sci. 16, 301–305.
Belmont, L. D. and Mitchison, T. J. (1996) Identification of a protein that interacts with tubulin dimers and increases the catastrophe rate of microtubules. Cell 84, 623–631.
Rubin, C. I. and Atweh, G. F. (2004) The role of stathmin in the regulation of the cell cycle. J. Cell. Biochem. 93, 242–250.
Cassimeris, L. (2002) The oncoprotein 18/stathmin family of microtubule destabilizers. Curr. Opin. Cell Biol. 14, 18–24.
Curmi, P. A., Gavet, O., Charbaut, E., et al. (1999) Stathmin and its phosphoprotein family: general properties, biochemical and functional interaction with tubulin. Cell Struct. Funct. 24, 345–357.
Charbaut, E., Curmi, P. A., Ozon, S., Lachkar, S., Redeker, V., and Sobel, A. (2001) Stathmin family proteins display specific molecular and tubulin binding properties. J. Biol. Chem. 276, 16,146–16,154.
Jourdain, L., Curmi, P., Sobel, A., Pantaloni, D., and Carlier, M. F. (1997) Stathmin: a tubulin-sequestering protein which forms a ternary T2S complex with two tubulin molecules. Biochemistry 36, 10,817–10,821.
Steinmetz, M. O., Kammerer, R. A., Jahnke, W., Goldie, K. N., Lustig, A., and van Oostrum, J. (2000) Op18/stathmin caps a kinked protofilament-like tubulin tetramer. EMBO J. 19, 572–580.
Gigant, B., Curmi, P. A., Martin-Barbey, C., et al. (2000) The 4 A X-ray structure of a tubulin:stathmin-like domain complex. Cell 102, 809–816.
Krouglova, T., Amayed, P., Engelborghs, Y., and Carlier, M. F. (2003) Fluorescence correlation spectroscopy analysis of the dynamics of tubulin interaction with RB3, a stathmin family protein. FEBS Lett. 546, 365–368.
Beeckmans, S. (1999) Chromatographic methods to study protein-protein interactions. Methods 19, 278–305.
Amayed, P., Pantaloni, D., and Carlier, M. F. (2002) The effect of stathmin phosphorylation on microtubule assembly depends on tubulin critical concentration. J. Biol. Chem. 277, 22,718–22,724.
Yarbrough, L. R. and Fishback, J. L. (1985) Kinetics of interaction of 2-amino-6-mercapto-9-beta-ribofuranosylpurine 5′-triphosphate with bovine brain tubulin. Biochemistry 24, 1708–1714.
Amayed, P., Carlier, M. F., and Pantaloni, D. (2000) Stathmin slows down guanosine diphosphate dissociation from tubulin in a phosphorylation-controlled fashion. Biochemistry 39, 12,295–12,302.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Humana Press Inc.
About this protocol
Cite this protocol
Carlier, MF. (2007). Measurements of Stathmin-Tubulin Interaction in Solution. In: Zhou, J. (eds) Microtubule Protocols. Methods in Molecular Medicine™, vol 137. Humana Press. https://doi.org/10.1007/978-1-59745-442-1_7
Download citation
DOI: https://doi.org/10.1007/978-1-59745-442-1_7
Publisher Name: Humana Press
Print ISBN: 978-1-58829-642-9
Online ISBN: 978-1-59745-442-1
eBook Packages: Springer Protocols