National Institutes of Health/National Institute of General Medical Sciences (NIH/NIGMS)
R35GM122510
米国
引用
ジャーナル: Nat Microbiol / 年: 2019 タイトル: An extensively glycosylated archaeal pilus survives extreme conditions. 著者: Fengbin Wang / Virginija Cvirkaite-Krupovic / Mark A B Kreutzberger / Zhangli Su / Guilherme A P de Oliveira / Tomasz Osinski / Nicholas Sherman / Frank DiMaio / Joseph S Wall / David ...著者: Fengbin Wang / Virginija Cvirkaite-Krupovic / Mark A B Kreutzberger / Zhangli Su / Guilherme A P de Oliveira / Tomasz Osinski / Nicholas Sherman / Frank DiMaio / Joseph S Wall / David Prangishvili / Mart Krupovic / Edward H Egelman / 要旨: Pili on the surface of Sulfolobus islandicus are used for many functions, and serve as receptors for certain archaeal viruses. The cells grow optimally at pH 3 and ~80 °C, exposing these ...Pili on the surface of Sulfolobus islandicus are used for many functions, and serve as receptors for certain archaeal viruses. The cells grow optimally at pH 3 and ~80 °C, exposing these extracellular appendages to a very harsh environment. The pili, when removed from cells, resist digestion by trypsin or pepsin, and survive boiling in sodium dodecyl sulfate or 5 M guanidine hydrochloride. We used electron cryo-microscopy to determine the structure of these filaments at 4.1 Å resolution. An atomic model was built by combining the electron density map with bioinformatics without previous knowledge of the pilin sequence-an approach that should prove useful for assemblies where all of the components are not known. The atomic structure of the pilus was unusual, with almost one-third of the residues being either threonine or serine, and with many hydrophobic surface residues. While the map showed extra density consistent with glycosylation for only three residues, mass measurements suggested extensive glycosylation. We propose that this extensive glycosylation renders these filaments soluble and provides the remarkable structural stability. We also show that the overall fold of the archaeal pilin is remarkably similar to that of archaeal flagellin, establishing common evolutionary origins.
平均露光時間: 2 sec. / 電子線照射量: 48 e/Å2 / 検出モード: INTEGRATING フィルム・検出器のモデル: FEI FALCON III (4k x 4k) 詳細: Images were stored containing 24 fractions, where each fraction corresponded to a dose of ~2 electrons per Angstrom^2.
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解析
ソフトウェア
名称: PHENIX / バージョン: dev_2919: / 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
Gctf
CTF補正
12
SPIDER
3次元再構成
13
RELION
3次元再構成
14
Rosetta
モデル精密化
15
PHENIX
モデル精密化
16
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: 104.97 ° / 軸方向距離/サブユニット: 4.94 Å / らせん対称軸の対称性: C1
3次元再構成
解像度: 4.1 Å / 解像度の算出法: OTHER / 粒子像の数: 181070 / 詳細: MODEL:MAP FSC, D99 / 対称性のタイプ: HELICAL