Purification of Mammalian Tubulins and Tubulin-Associated Proteins Using a P2A-Based Expression System
The microtubule cytoskeleton plays a crucial role in a myriad of cellular events, including mitosis, cell differentiation, migration, and the maintenance of cell shape. Microtubules are assembled from α- and β-tubulin heterodimers, whose biosynthesis is a complex process requiring the balanced production of α- and β-tubulin subunits. This chapter focuses on a method for the combined expression of tagged α- and β-tubulin dimers, their purification, and the isolation of co-purifying tubulin-associated proteins (TAPs) in mammalian cells. This approach is currently used in our laboratory to study tubulin function and to identify and characterize TAPs.
|Keywords||Affinity-based purification, Microtubule, Transient transfection, Tubulin, Tubulin-associated proteins|
|Persistent URL||dx.doi.org/10.1007/978-1-0716-0219-5_1, hdl.handle.net/1765/123395|
|Journal||Methods in Molecular Biology|
Yu, N. (Nuo), & Galjart, N.J. (2020). Purification of Mammalian Tubulins and Tubulin-Associated Proteins Using a P2A-Based Expression System. Methods in Molecular Biology, 2101, 1–17. doi:10.1007/978-1-0716-0219-5_1