Detection of Human βV-Tubulin Expression in Epithelial Cancer Cell Lines by Tubulin Proteomics.

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. βV-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. [ABSTRACT FROM AUTHOR]