9F3S

13pf mosaic 20%E254Q - 80% E254N microtubule from recombinant human tubulin decorated with EB3

9F3S の概要

• エントリーDOI: 10.2210/pdb9f3s/pdb
• 関連するPDBエントリー: 9F3B
• EMDBエントリー: 50178
• 分子名称: Microtubule-associated protein RP/EB family member 3, Detyrosinated tubulin alpha-1B chain, Tubulin beta-3 chain, ... (5 entities in total)
• 機能のキーワード: microtubule tubulin gtp cp cell cycle, structural protein
• 由来する生物種: Homo sapiens (human); 詳細
• タンパク質・核酸の鎖数: 14
• 化学式量合計: 649310.93

構造登録者

Estevez-Gallego, J.,Blum, T.B.,Steinmetz, M.O.,Surrey, T. (登録日: 2024-04-25, 公開日: 2025-03-19)

主引用文献

Estevez-Gallego, J.,Blum, T.B.,Ruhnow, F.,Gili, M.,Speroni, S.,Garcia-Castellanos, R.,Steinmetz, M.O.,Surrey, T.
Hydrolysis-deficient mosaic microtubules as faithful mimics of the GTP cap.
Nat Commun, 16:2396-2396, 2025

PubMed Abstract: A critical feature of microtubules is their GTP cap, a stabilizing GTP-tubulin rich region at growing microtubule ends. Microtubules polymerized in the presence of GTP analogs or from GTP hydrolysis-deficient tubulin mutants have been used as GTP-cap mimics for structural and biochemical studies. However, these analogs and mutants generate microtubules with diverse biochemical properties and lattice structures, leaving it unclear what is the most faithful GTP mimic and hence the structure of the GTP cap. Here, we generate a hydrolysis-deficient human tubulin mutant, αE254Q, with the smallest possible modification. We show that αE254Q-microtubules are stable, but still exhibit mild mutation-induced growth abnormalities. However, mixing two GTP hydrolysis-deficient tubulin mutants, αE254Q and αE254N, at an optimized ratio eliminates growth and lattice abnormalities, indicating that these 'mosaic microtubules' are faithful GTP cap mimics. Their cryo-electron microscopy structure reveals that longitudinal lattice expansion, but not protofilament twist, is the primary structural feature distinguishing the GTP-tubulin containing cap from the GDP-tubulin containing microtubule shaft. However, alterations in protofilament twist may be transiently needed to allow lattice compaction and GTP hydrolysis. Together, our results provide insights into the structural origin of GTP cap stability, the pathway of GTP hydrolysis and hence microtubule dynamic instability.

PubMed: 40064882
DOI: 10.1038/s41467-025-57555-6

実験手法

ELECTRON MICROSCOPY (4.2 Å)