ジャーナル: Proc Natl Acad Sci U S A / 年: 2019 タイトル: Effects of α-tubulin acetylation on microtubule structure and stability. 著者: Lisa Eshun-Wilson / Rui Zhang / Didier Portran / Maxence V Nachury / Daniel B Toso / Thomas Löhr / Michele Vendruscolo / Massimiliano Bonomi / James S Fraser / Eva Nogales / 要旨: Acetylation of K40 in α-tubulin is the sole posttranslational modification to mark the luminal surface of microtubules. It is still controversial whether its relationship with microtubule ...Acetylation of K40 in α-tubulin is the sole posttranslational modification to mark the luminal surface of microtubules. It is still controversial whether its relationship with microtubule stabilization is correlative or causative. We have obtained high-resolution cryo-electron microscopy (cryo-EM) reconstructions of pure samples of αTAT1-acetylated and SIRT2-deacetylated microtubules to visualize the structural consequences of this modification and reveal its potential for influencing the larger assembly properties of microtubules. We modeled the conformational ensembles of the unmodified and acetylated states by using the experimental cryo-EM density as a structural restraint in molecular dynamics simulations. We found that acetylation alters the conformational landscape of the flexible loop that contains αK40. Modification of αK40 reduces the disorder of the loop and restricts the states that it samples. We propose that the change in conformational sampling that we describe, at a location very close to the lateral contacts site, is likely to affect microtubule stability and function.
凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 310.15 K / 装置: FEI VITROBOT MARK IV / 詳細: Blotted for 4 seconds at blot force 10..
詳細
Acetylated and deacetylated tubulin preparations were produced by treating purified brain tubulin with the acetyltransferase TAT1 or the tubulin deacetylatase SIRT2 as done in Portran et al. Nat. Cell. Bio. 2017.
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
温度
最低: 77.0 K / 最高: 77.0 K
特殊光学系
エネルギーフィルター - 名称: GIF Quantum LS / エネルギーフィルター - スリット幅: 143 eV
詳細
Preliminary grid screening was performed manually and all of the alignments were initially done using a gold calibration grid.