Detection of human βV-tubulin expression in epithelial cancer cell lines by tubulin proteomics

Pascal Verdier-Pinard, Shohreh Shahabi, Fang Wang, Berta Burd, Hui Xiao, Gary L. Goldberg, George A. Orr, Susan Band Horwitz

Research output: Contribution to journalArticle

34 Scopus citations

Abstract

Tubulin, the constitutive protein of microtubules, is a heterodimeric protein with an α and β subunit, encoded in vertebrates by six and seven different genes, respectively. Each tubulin isotype can be identified by its divergent C-terminal sequence. Nevertheless, two groups of β-tubulin isotypes can be distinguished by sequence alignment; one includes βI-, βII-, βIVa-, and βIVb-tubulin, and the other includes βIII-, βV-, and βVI-tubulin. βIII-tubulin overexpression has been associated with microtubule destabilization and resistance to Taxol. Recent data indicate that mouse βV-tubulin overexpression in CHO cells results in profound microtubule disorganization and dependence of cells on Taxol for growth. Mouse and human βV-tubulin sequences display several differences, such as their respective extreme C-terminus, suggesting that they may have different effects on microtubule stability and different affinities for drugs. When high-resolution isoelectric focusing, in-gel CNBr cleavage, and mass spectrometry were combined, we detected for the first time the βV-tubulin protein in human cell lines and found that it was highly expressed in Hey, an epithelial ovarian cancer cell line. Our data confirm that human and rodent βV-tubulins are distinct and indicate that, regardless of species, βIII- and βV-tubulin may be expressed in a complementary pattern at the protein level. Therefore, both βIII- and βV-tubulin expression levels should be systematically determined to assess the role of differential tubulin isotype expression in the response of tumors to drugs targeting microtubules.

Original languageEnglish (US)
Pages (from-to)15858-15870
Number of pages13
JournalBiochemistry
Volume44
Issue number48
DOIs
StatePublished - Dec 6 2005

ASJC Scopus subject areas

  • Biochemistry

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    Verdier-Pinard, P., Shahabi, S., Wang, F., Burd, B., Xiao, H., Goldberg, G. L., Orr, G. A., & Horwitz, S. B. (2005). Detection of human βV-tubulin expression in epithelial cancer cell lines by tubulin proteomics. Biochemistry, 44(48), 15858-15870. https://doi.org/10.1021/bi051004p