ジャーナル: Mol Microbiol / 年: 2017 タイトル: Structural characterization of the NAP; the major adhesion complex of the human pathogen Mycoplasma genitalium. 著者: Margot P Scheffer / Luis Gonzalez-Gonzalez / Anja Seybert / Mercè Ratera / Michael Kunz / José M Valpuesta / Ignacio Fita / Enrique Querol / Jaume Piñol / Jaime Martín-Benito / Achilleas S Frangakis / 要旨: Mycoplasma genitalium, the causative agent of non-gonococcal urethritis and pelvic inflammatory disease in humans, is a small eubacterium that lacks a peptidoglycan cell wall. On the surface of its ...Mycoplasma genitalium, the causative agent of non-gonococcal urethritis and pelvic inflammatory disease in humans, is a small eubacterium that lacks a peptidoglycan cell wall. On the surface of its plasma membrane is the major surface adhesion complex, known as NAP that is essential for adhesion and gliding motility of the organism. Here, we have performed cryo-electron tomography of intact cells and detergent permeabilized M. genitalium cell aggregates, providing sub-tomogram averages of free and cell-attached NAPs respectively, revealing a tetrameric complex with two-fold rotational (C2) symmetry. Each NAP has two pairs of globular lobes (named α and β lobes), arranged as a dimer of heterodimers with each lobe connected by a stalk to the cell membrane. The β lobes are larger than the α lobes by 20%. Classification of NAPs showed that the complex can tilt with respect to the cell membrane. A protein complex containing exclusively the proteins P140 and P110, was purified from M. genitalium and was structurally characterized by negative-stain single particle EM reconstruction. The close structural similarity found between intact NAPs and the isolated P140/P110 complexes, shows that dimers of P140/P110 heterodimers are the only components of the extracellular region of intact NAPs in M. genitalium.
ダウンロード / ファイル: emd_3753.map.gz / 形式: CCP4 / 大きさ: 1 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Negative stain 3D single particle reconstruction of the isolated surface adhesin P140/P110 complex of Mycoplasma genitalium.
ボクセルのサイズ
X=Y=Z: 4.32 Å
密度
表面レベル
登録者による: 0.1047 / ムービー #1: 0.1047
最小 - 最大
-0.16509591 - 0.3144459
平均 (標準偏差)
-0.000034514454 (±0.031377893)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
64
64
64
Spacing
64
64
64
セル
A=B=C: 276.48 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
4.32
4.32
4.32
M x/y/z
64
64
64
origin x/y/z
0.000
0.000
0.000
length x/y/z
276.480
276.480
276.480
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
64
64
64
D min/max/mean
-0.165
0.314
-0.000
-
添付データ
-
試料の構成要素
-
全体 : Negative stain 3D single particle reconstruction of the isolated ...
全体
名称: Negative stain 3D single particle reconstruction of the isolated surface adhesin P140/P110 complex of Mycoplasma genitalium.
要素
複合体: Negative stain 3D single particle reconstruction of the isolated surface adhesin P140/P110 complex of Mycoplasma genitalium.
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超分子 #1: Negative stain 3D single particle reconstruction of the isolated ...
超分子
名称: Negative stain 3D single particle reconstruction of the isolated surface adhesin P140/P110 complex of Mycoplasma genitalium. タイプ: complex / ID: 1 / 親要素: 0 詳細: NAP complexes were extracted from the cell membranes of a modified M. genitalium G37 strain by using the detergent n-octyl-beta-D-glucopyranoside, where gene mg192 had been extended to ...詳細: NAP complexes were extracted from the cell membranes of a modified M. genitalium G37 strain by using the detergent n-octyl-beta-D-glucopyranoside, where gene mg192 had been extended to include a region coding for a six-histidines tag at the C-terminus end of the P110 protein, the gene product of mg192.
由来(天然)
生物種: Mycoplasma genitalium G37 (バクテリア)
組換発現
生物種: Mycoplasma genitalium G37 (バクテリア)
分子量
実験値: 495 kDa/nm
-
実験情報
-
構造解析
手法
ネガティブ染色法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
緩衝液
pH: 7.4
染色
タイプ: NEGATIVE / 材質: Uranyl Acetate
-
電子顕微鏡法
顕微鏡
FEI TECNAI F20
撮影
フィルム・検出器のモデル: FEI EAGLE (4k x 4k) / 平均電子線量: 100.0 e/Å2