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Title | Tubulin engineering by semi-synthesis reveals that polyglutamylation directs detyrosination. |
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Journal, issue, pages | Nat Chem, Vol. 15, Issue 8, Page 1179-1187, Year 2023 |
Publish date | Jun 29, 2023 |
Authors | Eduard Ebberink / Simon Fernandes / Georgios Hatzopoulos / Ninad Agashe / Po-Han Chang / Nora Guidotti / Timothy M Reichart / Luc Reymond / Marie-Claire Velluz / Fabian Schneider / Cédric Pourroy / Carsten Janke / Pierre Gönczy / Beat Fierz / Charlotte Aumeier / |
PubMed Abstract | Microtubules, a critical component of the cytoskeleton, carry post-translational modifications (PTMs) that are important for the regulation of key cellular processes. Long-lived microtubules, in ...Microtubules, a critical component of the cytoskeleton, carry post-translational modifications (PTMs) that are important for the regulation of key cellular processes. Long-lived microtubules, in neurons particularly, exhibit both detyrosination of α-tubulin and polyglutamylation. Dysregulation of these PTMs can result in developmental defects and neurodegeneration. Owing to a lack of tools to study the regulation and function of these PTMs, the mechanisms that govern such PTM patterns are not well understood. Here we produce fully functional tubulin carrying precisely defined PTMs within its C-terminal tail. We ligate synthetic α-tubulin tails-which are site-specifically glutamylated-to recombinant human tubulin heterodimers by applying a sortase- and intein-mediated tandem transamidation strategy. Using microtubules reconstituted with these designer tubulins, we find that α-tubulin polyglutamylation promotes its detyrosination by enhancing the activity of the tubulin tyrosine carboxypeptidase vasohibin/small vasohibin-binding protein in a manner dependent on the length of polyglutamyl chains. We also find that modulating polyglutamylation levels in cells results in corresponding changes in detyrosination, corroborating the link between the detyrosination cycle to polyglutamylation. |
External links | Nat Chem / PubMed:37386282 |
Methods | EM (helical sym.) |
Resolution | 3.68 - 3.79 Å |
Structure data | EMDB-15118: 13pf undecorated microtubule from recombinant human tubulin (alpha1B, beta3) lacking the C-terminal tail EMDB-15119: 13pf undecorated microtubule from recombinant human tubulin (alpha1B, beta3) with spliced unmodified C-terminal tail on alpha1B. EMDB-15120: 13pf undecorated microtubule from recombinant human tubulin (alpha1B, beta3) with spliced C-terminal tail containing 10E branch on alpha1B. |
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