Related Experiment Video

Updated: Sep 9, 2025

Extracting Modified Microtubules from Mammalian Cells to Study Microtubule-Protein Complexes by Cryo-Electron Microscopy
08:02

Extracting Modified Microtubules from Mammalian Cells to Study Microtubule-Protein Complexes by Cryo-Electron Microscopy

Published on: March 3, 2023

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MATCAP1 preferentially binds an expanded tubulin conformation to generate detyrosinated and ΔC2 α-tubulin

Yang Yue1, Takashi Hotta1, Ryoma Ohi1

  • 1Department of Cell & Developmental Biology, University of Michigan Medical School, Ann Arbor, MI, USA. 48109.

Biorxiv : the Preprint Server for Biology
|September 2, 2025
PubMed

Insights

Metallopeptidase MATCAP1 modifies alpha-tubulin via detyrosination (ΔY) and ΔC2. Microtubule lattice conformation dictates MATCAP1 binding and activity, controlling these posttranslational modifications (PTMs).

Area of Science:

  • Cell Biology
  • Biochemistry
  • Structural Biology

Background:

  • Microtubules are essential cytoskeletal polymers involved in cell division, motility, and intracellular transport.
  • Posttranslational modifications (PTMs) of alpha-tubulin, such as detyrosination (ΔY) and ΔC2, regulate microtubule interactions with associated proteins and motors.
  • The mechanisms generating specific PTM patterns on microtubules remain largely unknown.

Purpose of the Study:

  • To investigate the in vitro microtubule binding behavior of metallopeptidase MATCAP1.
  • To elucidate the mechanism by which MATCAP1 generates ΔY and ΔC2 modifications on alpha-tubulin.

Main Methods:

  • In vitro reconstitution assays using purified proteins.
  • Analysis of MATCAP1 binding to microtubules under various conditions (e.g., altered tubulin conformation).
  • Biochemical assays to detect α-tubulin detyrosination.

Main Results:

  • MATCAP1 preferentially binds to microtubules with expanded tubulin conformations.
  • Expanded microtubule lattices are induced by preventing β-tubulin GTP hydrolysis, Taxol treatment, or kinesin-1 activity.
  • MATCAP1 sequentially removes tyrosine (generating ΔY) and glutamate (generating ΔC2) residues from α-tubulin C-terminus.
  • MATCAP1 exhibits long dwell times on expanded microtubule lattices.

Conclusions:

  • Microtubule lattice conformation is a critical determinant for MATCAP1 binding and enzymatic activity.
  • The study reveals a mechanism linking microtubule structure to the generation of specific α-tubulin PTMs.
  • Understanding MATCAP1's activity provides insights into the regulation of microtubule function through PTMs.
Keywords:
MATCAP1Microtubuledetyrosinationmicrotubule conformationposttranslational modificationtubulinΔC2 modification

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