Preparation of Pure Tyrosinated or Detyrosinated Tubulin Isoforms

Alpha tubulin comprises a C-terminal tyrosine residue, which is subject to cyclic removal from the peptide chain by a still uncharacterized carboxypeptidase and re-addition to the chain by a tubulin tyrosine ligase. We have shown in different animal or human models that the presence or absence of the tyrosine residue had dramatic consequences for both tumor progression and neuronal organization. In cells, tubulin detyrosination impairs the proper localization of CAP-Gly proteins at microtubule + end, compromises the activity of microtubule-depolymerizing motors of the Kinesin 13 family, and favors both spastin microtubule-severing activity and kinesin 1 processivity. The biochemical basis for these cellular effects of tubulin detyrosination can now be investigated in reconstituted systems in vitro using homogeneous solutions of polymerizable tyrosinated or detyrosinated tubulin.