National Institutes of Health/National Institute Of Allergy and Infectious Diseases (NIH/NIAID)
AI114902
米国
Canadian Institutes of Health Research (CIHR)
MOP125959
カナダ
French National Research Agency
ANR14CE14001002
フランス
Fondation pour la Recherche Medicale
フランス
引用
ジャーナル: Structure / 年: 2017 タイトル: Cryoelectron Microscopy Reconstructions of the Pseudomonas aeruginosa and Neisseria gonorrhoeae Type IV Pili at Sub-nanometer Resolution. 著者: Fengbin Wang / Mathieu Coureuil / Tomasz Osinski / Albina Orlova / Tuba Altindal / Gaël Gesbert / Xavier Nassif / Edward H Egelman / Lisa Craig / 要旨: We report here cryoelectron microscopy reconstructions of type IV pili (T4P) from two important human pathogens, Pseudomonas aeruginosa and Neisseria gonorrhoeae, at ∼ 8 and 5 Å resolution, ...We report here cryoelectron microscopy reconstructions of type IV pili (T4P) from two important human pathogens, Pseudomonas aeruginosa and Neisseria gonorrhoeae, at ∼ 8 and 5 Å resolution, respectively. The two structures reveal distinct arrangements of the pilin globular domains on the pilus surfaces, which impart different helical parameters, but similar packing of the conserved N-terminal α helices, α1, in the filament core. In contrast to the continuous α helix seen in the X-ray crystal structures of the P. aeruginosa and N. gonorrhoeae pilin subunits, α1 in the pilus filaments has a melted segment located between conserved helix-breaking residues Gly14 and Pro22, as seen for the Neisseria meningitidis T4P. Using mutagenesis we show that Pro22 is critical for pilus assembly, as are Thr2 and Glu5, which are positioned to interact in the hydrophobic filament core. These structures provide a framework for understanding T4P assembly, function, and biophysical properties.
A: Fimbrial protein B: Fimbrial protein C: Fimbrial protein D: Fimbrial protein E: Fimbrial protein F: Fimbrial protein G: Fimbrial protein H: Fimbrial protein I: Fimbrial protein J: Fimbrial protein K: Fimbrial protein L: Fimbrial protein M: Fimbrial protein N: Fimbrial protein O: Fimbrial protein P: Fimbrial protein Q: Fimbrial protein R: Fimbrial protein S: Fimbrial protein T: Fimbrial protein U: Fimbrial protein
平均露光時間: 2 sec. / 電子線照射量: 20 e/Å2 / 検出モード: INTEGRATING フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 詳細: Images were stored containing seven parts, where each part represented a set of frames corresponding to a dose of ~20 electrons per Angstrom^2. The full dose image stack was used for the ...詳細: Images were stored containing seven parts, where each part represented a set of frames corresponding to a dose of ~20 electrons per Angstrom^2. The full dose image stack was used for the estimation of the CTF as well as for boxing filaments. Only the first two parts were used for the reconstruction (~5 electrons per Angstrom^2)
画像スキャン
動画フレーム数/画像: 7
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解析
ソフトウェア
名称: PHENIX / バージョン: dev_2471: / 分類: 精密化
EMソフトウェア
ID
名称
カテゴリ
1
EMAN2
粒子像選択
2
EPU
画像取得
4
CTFFIND3
CTF補正
7
Rosetta
モデルフィッティング
8
UCSF Chimera
モデルフィッティング
10
SPIDER
初期オイラー角割当
11
SPIDER
最終オイラー角割当
13
SPIDER
3次元再構成
14
PHENIX
モデル精密化
15
Coot
モデル精密化
CTF補正
タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION
らせん対称
回転角度/サブユニット: 87.3 ° / 軸方向距離/サブユニット: 10.5 Å / らせん対称軸の対称性: C1
3次元再構成
解像度: 8 Å / 解像度の算出法: OTHER / 粒子像の数: 31231 / 詳細: model-map FSC 0.38 cut-off / 対称性のタイプ: HELICAL