Elsevier

Analytical Biochemistry

Volume 384, Issue 2, 15 January 2009, Pages 197-206
Analytical Biochemistry

Review
Tubulin proteomics: Towards breaking the code

This review is dedicated to the memory of our esteemed colleague Professor George A. Orr, whose ideas formed the foundation of this research. His creativity and dedication continue to inspire our work in tubulin proteomics.
https://doi.org/10.1016/j.ab.2008.09.020Get rights and content

Section snippets

Tubulin isotypes and functional significance: first hints of a tubulin code

Tubulin is a 100 kDa heterodimer formed by an α- and a β-subunit that are equivalent in size and structure [14]. In mammals, tubulin represents about 3–4% of the total proteins in cells and up to 10% in brain. In humans, eight α-tubulin and seven β-tubulin genes have been identified [15], [16]. These tubulin isotypes have been detected at the mRNA and/or protein level and are differentially distributed in tissues [16]. Additional complexity is generated by the fact that each isotype can undergo

Analysis of tubulin expression profiles: key contributions from mass spectrometry

Tubulin isotype expression profiling has been frequently assessed by RT-PCR of mRNA, and at the protein level by antibody-based approaches [2]. These approaches are useful in a high-throughput setting, but results must be interpreted cautiously. This is particularly relevant to β-tubulin mRNA, because it appears to be autoregulated by the level of the free β-tubulin in cells [36], implying that levels of mRNA may not reflect levels of the corresponding tubulin. Antibodies directed against the

Use and development of tubulin proteomics

Collectively, high-resolution IEF-MS and LC-MS determination of pI and mass of human tubulin isotypes, respectively, constitute a bi-dimensional analysis that confirms which sequences are expressed at the protein level. Nevertheless, each of these approaches has its limitations, and some tubulin species may be present below the level of detection. Taxol-dependent polymerization does not enrich for specific β-tubulin isotypes selectively [62], but it may exclude from the analysis a

Acknowledgments

This work was supported by ANR-05-BLAN-SPV00551 (to D.L) and INCa-Cancéropôle PACA 2003 (Plateforme Protéomique Timone; to D.B) and NIH grants CA077263 and CA124898, and the National Foundation for Cancer Research (to S.B.H.) and CA110150 (to R.H.A). E. P. is supported by a Cancer Institute New South Wales Early Career Development Fellowship. L.M.M. is supported by an NIH postdoctoral fellowship (CA125923).

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    Present address: Children’s Cancer Institute Australia for Medical Research, Pharmacoproteomics Program, P.O. Box 81 (High Street), Randwick, NSW 2031, Australia.

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