Methods in tubulin proteomics

Leah M. Miller, Hui Xiao, Berta Burd, Susan Band Horwitz, Ruth Hogue Angeletti, Pascal Verdier-Pinard

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

New analytical methods are needed for the successful outcome of experiments aimed at characterizing mechanisms of microtubule dynamics and at understanding the effects of drugs on microtubules. The identification of tubulin isotypes and of regions of the microtubule involved in drug interactions has been advanced by proteomic methodologies. The diversity of tubulin sequences and posttranslational modifications (PTMs) can generate a complex mixture of heterodimers with unique molecular dynamics driving specific functions. Mass spectrometry (MS)-based approaches have been developed, and in combination with chromatographic and/or electrophoretic separation of tubulin polypeptides or peptides, they have contributed to our understanding of tubulin proteomics. We present protocols that we have used for the analysis of tubulin isotypes and PTMs present in tubulin isolated from cells in culture or tissues and for the identification of tubulin regions altered by microtubule-stabilizing agents. Tubulin proteomics complements structural and computer modeling information for a high-resolution view of microtubule dynamics and its alteration by drugs. These methodologies will help in providing insights into tubulin isotype-specific functions and in the design of drugs targeting either all tubulin heterodimers indiscriminately or only those containing specific isotypes.

Original languageEnglish (US)
Pages (from-to)105-126
Number of pages22
JournalMethods in cell biology
Volume95
Issue numberC
DOIs
StatePublished - 2010

ASJC Scopus subject areas

  • Cell Biology

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    Miller, L. M., Xiao, H., Burd, B., Horwitz, S. B., Angeletti, R. H., & Verdier-Pinard, P. (2010). Methods in tubulin proteomics. Methods in cell biology, 95(C), 105-126. https://doi.org/10.1016/S0091-679X(10)95007-3