title: Tyrosination of microtubules and non-assembled tubulin in brain slices.
abstract: Brain slices were used to examine comparatively the incorporation of [14C]tyrosine into the C terminus of alpha-tubulin of the microtubule and non-assembled tubulin pools. We found that the incorporation of [14C]tyrosine from 5 min up to 60 min of incubation was higher in microtubules than in non-assembled tubulin. The possibility that this result was due to the activity of tubulin carboxypeptidase or tubulin:tyrosine ligase during the in vitro isolation of tubulin was discarded. We also found that tubulin:tyrosine ligase was mainly associated with microtubules when brain slices were homogenized under microtubule-preserving conditions. Conversely the enzyme behaved as a soluble entity when homogenization was performed under conditions that do not preserve microtubules. In addition, soluble tubulin:tyrosine ligase did not become sedimentable when in vitro conditions were changed to induce the formation of microtubules. The results presented in this work indicate the possibility that, in vivo, microtubules and not tubulin dimers are the major substrate for tubulin:tyrosine ligase. This is in contrast with previous findings from in vitro experiments, which showed a preference of the ligase for non-assembled tubulin.
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism*; D002268:Carboxypeptidases / Q000378:metabolism; D066298:In Vitro Techniques; D008870:Microtubules / Q000378:metabolism*; D010453:Peptide Synthases / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D014404:Tubulin / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D014404:Tubulin; D014443:Tyrosine; D002268:Carboxypeptidases; C023212:tyrosyltubulin carboxypeptidase; D010453:Peptide Synthases; C023213:tyrosyltubulin ligase