ジャーナル: Elife / 年: 2013 タイトル: In situ structural analysis of the Yersinia enterocolitica injectisome. 著者: Mikhail Kudryashev / Marco Stenta / Stefan Schmelz / Marlise Amstutz / Ulrich Wiesand / Daniel Castaño-Díez / Matteo T Degiacomi / Stefan Münnich / Christopher Ke Bleck / Julia Kowal / ...著者: Mikhail Kudryashev / Marco Stenta / Stefan Schmelz / Marlise Amstutz / Ulrich Wiesand / Daniel Castaño-Díez / Matteo T Degiacomi / Stefan Münnich / Christopher Ke Bleck / Julia Kowal / Andreas Diepold / Dirk W Heinz / Matteo Dal Peraro / Guy R Cornelis / Henning Stahlberg / 要旨: Injectisomes are multi-protein transmembrane machines allowing pathogenic bacteria to inject effector proteins into eukaryotic host cells, a process called type III secretion. Here we present the ...Injectisomes are multi-protein transmembrane machines allowing pathogenic bacteria to inject effector proteins into eukaryotic host cells, a process called type III secretion. Here we present the first three-dimensional structure of Yersinia enterocolitica and Shigella flexneri injectisomes in situ and the first structural analysis of the Yersinia injectisome. Unexpectedly, basal bodies of injectisomes inside the bacterial cells showed length variations of 20%. The in situ structures of the Y. enterocolitica and S. flexneri injectisomes had similar dimensions and were significantly longer than the isolated structures of related injectisomes. The crystal structure of the inner membrane injectisome component YscD appeared elongated compared to a homologous protein, and molecular dynamics simulations documented its elongation elasticity. The ring-shaped secretin YscC at the outer membrane was stretched by 30-40% in situ, compared to its isolated liposome-embedded conformation. We suggest that elasticity is critical for some two-membrane spanning protein complexes to cope with variations in the intermembrane distance. DOI:http://dx.doi.org/10.7554/eLife.00792.001.
全体 : Yersinia enterocolitica intact bacterial cells containing type II...
全体
名称: Yersinia enterocolitica intact bacterial cells containing type III secretion injectisomes
要素
試料: Yersinia enterocolitica intact bacterial cells containing type III secretion injectisomes
タンパク質・ペプチド: Bacterial Type III Secretion System
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超分子 #1000: Yersinia enterocolitica intact bacterial cells containing type II...
超分子
名称: Yersinia enterocolitica intact bacterial cells containing type III secretion injectisomes タイプ: sample / ID: 1000 詳細: Bacteria were taken from cell culture and were rapidly prepared for cryo imaging 集合状態: various / Number unique components: 9
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分子 #1: Bacterial Type III Secretion System
分子
名称: Bacterial Type III Secretion System / タイプ: protein_or_peptide / ID: 1 / Name.synonym: Injectisome, T3SS / 組換発現: No / データベース: NCBI
由来(天然)
生物種: Yersinia enterocolitica (腸炎エルシニア)
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実験情報
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構造解析
手法
クライオ電子顕微鏡法
解析
サブトモグラム平均法
試料の集合状態
particle
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試料調製
緩衝液
詳細: PBS
グリッド
詳細: Quantifoil holey carbon grids with various hole sizes
凍結
凍結剤: ETHANE / チャンバー内温度: 77 K / 装置: FEI VITROBOT MARK IV
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
特殊光学系
エネルギーフィルター - 名称: GIF Tridem エネルギーフィルター - エネルギー下限: 0.0 eV エネルギーフィルター - エネルギー上限: 20.0 eV