National Institutes of Health/National Institute Of Allergy and Infectious Diseases
R01AI127401
米国
引用
ジャーナル: Nat Microbiol / 年: 2019 タイトル: In vivo structure of the Legionella type II secretion system by electron cryotomography. 著者: Debnath Ghosal / Ki Woo Kim / Huaixin Zheng / Mohammed Kaplan / Hilary K Truchan / Alberto E Lopez / Ian E McIntire / Joseph P Vogel / Nicholas P Cianciotto / Grant J Jensen / 要旨: The type II secretion system (T2SS) is a multiprotein envelope-spanning assembly that translocates a wide range of virulence factors, enzymes and effectors through the outer membrane of many Gram- ...The type II secretion system (T2SS) is a multiprotein envelope-spanning assembly that translocates a wide range of virulence factors, enzymes and effectors through the outer membrane of many Gram-negative bacteria. Here, using electron cryotomography and subtomogram averaging methods, we reveal the in vivo structure of an intact T2SS imaged within the human pathogen Legionella pneumophila. Although the T2SS has only limited sequence and component homology with the evolutionarily related type IV pilus (T4P) system, we show that their overall architectures are remarkably similar. Despite similarities, there are also differences, including, for example, that the T2SS-ATPase complex is usually present but disengaged from the inner membrane, the T2SS has a much longer periplasmic vestibule and it has a short-lived flexible pseudopilus. Placing atomic models of the components into our electron cryotomography map produced a complete architectural model of the intact T2SS that provides insights into the structure and function of its components, its position within the cell envelope and the interactions between its different subcomplexes.
A: 672.0 Å / B: 1007.99994 Å / C: 672.0 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
8.4
8.4
8.4
M x/y/z
80
120
80
origin x/y/z
0.000
0.000
0.000
length x/y/z
672.000
1008.000
672.000
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-200
-200
-200
NX/NY/NZ
401
401
401
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
80
120
80
D min/max/mean
0.000
1.000
0.312
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添付データ
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試料の構成要素
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全体 : Subtomogram average of the Legionella pneumophila type II secreti...
全体
名称: Subtomogram average of the Legionella pneumophila type II secretion system (aligning the OM-complex).
要素
複合体: Subtomogram average of the Legionella pneumophila type II secretion system (aligning the OM-complex).
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超分子 #1: Subtomogram average of the Legionella pneumophila type II secreti...
超分子
名称: Subtomogram average of the Legionella pneumophila type II secretion system (aligning the OM-complex). タイプ: complex / ID: 1 / 親要素: 0 詳細: We used electron cryotomography and subtomogram averaging to visualize L. pneumophila type II secretion system complex directly in intact, frozen-hydrated bacterial cells
由来(天然)
生物種: Legionella pneumophila (バクテリア) / 株: Lp02
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実験情報
-
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
cell
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試料調製
緩衝液
pH: 6.9
グリッド
材質: COPPER
凍結
凍結剤: ETHANE-PROPANE
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 平均電子線量: 1.2 e/Å2
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD
実験機器
モデル: Titan Krios / 画像提供: FEI Company
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画像解析
最終 再構成
想定した対称性 - 点群: C2 (2回回転対称) / 解像度のタイプ: BY AUTHOR / 解像度: 40.0 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: IMOD / 使用したサブトモグラム数: 440