ReviewTubulin proteomics: Towards breaking the code
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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2019, Biochimica et Biophysica Acta - Molecular Basis of DiseaseCitation Excerpt :This was determined by immunocytochemistry (using mouse antibodies for the various modifications being investigated), double-label immunofluorescence imaging (using inverted fluorescence and inverted laser-scanning confocal fluorescence microscopy), and western blotting. While the data is convincing, particularly due to the fact that it corroborates previously published results, advancements in mass spectrometry have been shown to be a robust means of characterizing and analyzing tubulin [87–89]. Additionally, it has been shown that SRM or MRM are more informative, primarily when it comes to quantification and highly complex sample analysis, than western blot analysis alone, although they may still be used in conjunction [90,91].
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2017, Biomedicine and PharmacotherapyDelineating the Role of βIV-Tubulins in Pancreatic Cancer: βIVb-Tubulin Inhibition Sensitizes Pancreatic Cancer Cells to Vinca Alkaloids
2016, Neoplasia (United States)Citation Excerpt :Dysregulation of proteins which comprise the cell cytoskeleton and/or microtubule network have been implicated in chemotherapy drug resistance and aggressive disease in different tumor types [7,8]. The microtubule network forms part of the cell cytoskeleton and consists of cylindrical assemblies of α- and β-tubulin heterodimers [8,9]. Microtubules are essential for spindle formation during chromosome alignment and segregation processes of mitosis [8,10,11].
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2015, Seminars in Pediatric NeurologyNeuronal process structure and growth proteins are targets of heavy PTM regulation during brain development
2014, Journal of Proteomics
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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.