EMDB/PDB/SASBDBから引用されている文献のデータベース

キーワード
構造解析手法	All X線回折 NMR 電子顕微鏡小角散乱中性子線回折線維回折粉末回折赤外分光 EPR FRET 理論モデルハイブリッド
著者・登録者
ジャーナル
IF	>30 >20 >10 >5 all

タイトル	Structural transitions in the GTP cap visualized by cryo-electron microscopy of catalytically inactive microtubules.
ジャーナル・号・ページ	Proc Natl Acad Sci U S A, Vol. 119, Issue 2, Year 2022
掲載日	2022年1月11日
著者	Benjamin J LaFrance / Johanna Roostalu / Gil Henkin / Basil J Greber / Rui Zhang / Davide Normanno / Chloe O McCollum / Thomas Surrey / Eva Nogales /
PubMed 要旨	Microtubules (MTs) are polymers of αβ-tubulin heterodimers that stochastically switch between growth and shrinkage phases. This dynamic instability is critically important for MT function. It is ...Microtubules (MTs) are polymers of αβ-tubulin heterodimers that stochastically switch between growth and shrinkage phases. This dynamic instability is critically important for MT function. It is believed that GTP hydrolysis within the MT lattice is accompanied by destabilizing conformational changes and that MT stability depends on a transiently existing GTP cap at the growing MT end. Here, we use cryo-electron microscopy and total internal reflection fluorescence microscopy of GTP hydrolysis-deficient MTs assembled from mutant recombinant human tubulin to investigate the structure of a GTP-bound MT lattice. We find that the GTP-MT lattice of two mutants in which the catalytically active glutamate in α-tubulin was substituted by inactive amino acids (E254A and E254N) is remarkably plastic. Undecorated E254A and E254N MTs with 13 protofilaments both have an expanded lattice but display opposite protofilament twists, making these lattices distinct from the compacted lattice of wild-type GDP-MTs. End-binding proteins of the EB family have the ability to compact both mutant GTP lattices and to stabilize a negative twist, suggesting that they promote this transition also in the GTP cap of wild-type MTs, thereby contributing to the maturation of the MT structure. We also find that the MT seam appears to be stabilized in mutant GTP-MTs and destabilized in GDP-MTs, supporting the proposal that the seam plays an important role in MT stability. Together, these structures of catalytically inactive MTs add mechanistic insight into the GTP state of MTs, the stability of the GTP- and GDP-bound lattice, and our overall understanding of MT dynamic instability.
リンク	Proc Natl Acad Sci U S A / PubMed:34996871 / PubMed Central
手法	EM (らせん対称)
解像度	3.4 - 5.0 Å
構造データ	25156 7sj7 EMDB-25156, PDB-7sj7: Undecorated 13pf wildtype microtubule from recombinant human tubulin 手法: EM (らせん対称) / 解像度: 3.8 Å 25157 7sj8 EMDB-25157, PDB-7sj8: 13pf wildtype microtubule from recombinant human tubulin decorated with kinesin 手法: EM (らせん対称) / 解像度: 3.6 Å EMDB-25158: Undecorated 13pf E254A microtubule from recombinant human tubulin 手法: EM (らせん対称) / 解像度: 3.4 Å 25159 7sj9 EMDB-25159, PDB-7sj9: 13pf E254A microtubule from recombinant human tubulin decorated with EB3 手法: EM (らせん対称) / 解像度: 3.8 Å 25160 7sja EMDB-25160, PDB-7sja: Undecorated 13pf E254N microtubule from recombinant human tubulin 手法: EM (らせん対称) / 解像度: 3.8 Å EMDB-25161: 13pf E254N microtubule from recombinant human tubulin decorated with EB3 手法: EM (らせん対称) / 解像度: 5.0 Å
化合物	ChemComp-GTP: GUANOSINE-5'-TRIPHOSPHATE / GTP / GTP, エネルギー貯蔵分子YM / グアノシン三リン酸 ChemComp-MG: Unknown entry / マグネシウムジカチオン ChemComp-GDP: GUANOSINE-5'-DIPHOSPHATE / GDP / GDP, エネルギー貯蔵分子YM / グアノシン二リン酸
由来	homo sapiens (ヒト)
キーワード	STRUCTURAL PROTEIN (タンパク質) / microtubule (微小管) / cell division (細胞分裂) / cytoskeleton (細胞骨格) / GTPase (GTPアーゼ)