Abstract
Hydrodynamic analysis is a powerful tool to dissect the molecular architecture of macromolecular protein assemblies. These techniques have been successfully used in yeast systems but are also well suited to the analysis of protein complexes from human cells. Furthermore, the combination of hydrodynamic analysis with siRNA mediated protein depletion provides an excellent system to probe the composition of protein complexes isolated from human cells. In this chapter we describe the use of these approaches in the analysis of macromolecular protein complexes during mitosis in human cells, using the kinetochore as an example.
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Acknowledgments
The authors would like to thank the members of the McAinsh lab for helpful discussions and reading of the manuscript. We would also like to thank Hiro Yamano for equipment and expertise relating to the glycerol gradient ultracentrifugation experiments. This work was supported by Marie Curie Cancer Care.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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McClelland, S.E., McAinsh, A.D. (2009). Hydrodynamic Analysis of Human Kinetochore Complexes During Mitosis. In: McAinsh, A. (eds) Mitosis. Methods in Molecular Biology, vol 545. Humana Press. https://doi.org/10.1007/978-1-60327-993-2_5
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DOI: https://doi.org/10.1007/978-1-60327-993-2_5
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