Reconstitution of the recombinant human γ-tubulin ring complex.

Martin Würtz, Anna Böhler, Annett Neuner, Erik Zupa, Lukas Rohland, Peng Liu, Bram J A Vermeulen, Stefan Pfeffer, Sebastian Eustermann, Elmar Schiebel

Open biology 2021 Feb

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Cryo-electron microscopy recently resolved the structure of the vertebrate γ-tubulin ring complex (γ-TuRC) purified from Xenopus laevis egg extract and human cells to near-atomic resolution. These studies clarified the arrangement and stoichiometry of γ-TuRC components and revealed that one molecule of actin and the small protein MZT1 are embedded into the complex. Based on this structural census of γ-TuRC core components, we developed a recombinant expression system for the reconstitution and purification of human γ-TuRC from insect cells. The recombinant γ-TuRC recapitulates the structure of purified native γ-TuRC and has similar functional properties in terms of microtubule nucleation and minus end capping. This recombinant system is a central step towards deciphering the activation mechanisms of the γ-TuRC and the function of individual γ-TuRC core components.

Citation

Martin Würtz, Anna Böhler, Annett Neuner, Erik Zupa, Lukas Rohland, Peng Liu, Bram J A Vermeulen, Stefan Pfeffer, Sebastian Eustermann, Elmar Schiebel. Reconstitution of the recombinant human γ-tubulin ring complex. Open biology. 2021 Feb;11(2):200325