ジャーナル: Science / 年: 2015 タイトル: DENGUE VIRUS. Cryo-EM structure of an antibody that neutralizes dengue virus type 2 by locking E protein dimers. 著者: Guntur Fibriansah / Kristie D Ibarra / Thiam-Seng Ng / Scott A Smith / Joanne L Tan / Xin-Ni Lim / Justin S G Ooi / Victor A Kostyuchenko / Jiaqi Wang / Aravinda M de Silva / Eva Harris / ...著者: Guntur Fibriansah / Kristie D Ibarra / Thiam-Seng Ng / Scott A Smith / Joanne L Tan / Xin-Ni Lim / Justin S G Ooi / Victor A Kostyuchenko / Jiaqi Wang / Aravinda M de Silva / Eva Harris / James E Crowe / Shee-Mei Lok / 要旨: There are four closely-related dengue virus (DENV) serotypes. Infection with one serotype generates antibodies that may cross-react and enhance infection with other serotypes in a secondary infection. ...There are four closely-related dengue virus (DENV) serotypes. Infection with one serotype generates antibodies that may cross-react and enhance infection with other serotypes in a secondary infection. We demonstrated that DENV serotype 2 (DENV2)-specific human monoclonal antibody (HMAb) 2D22 is therapeutic in a mouse model of antibody-enhanced severe dengue disease. We determined the cryo-electron microscopy (cryo-EM) structures of HMAb 2D22 complexed with two different DENV2 strains. HMAb 2D22 binds across viral envelope (E) proteins in the dimeric structure, which probably blocks the E protein reorganization required for virus fusion. HMAb 2D22 "locks" two-thirds of or all dimers on the virus surface, depending on the strain, but neutralizes these DENV2 strains with equal potency. The epitope defined by HMAb 2D22 is a potential target for vaccines and therapeutics.
ダウンロード / ファイル: emd_2997.map.gz / 形式: CCP4 / 大きさ: 500 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Reconstruction of Dengue virus serotype 2 strain New Guinea-C complexed with human antibody 2D22 Fab at 4 degrees C.
ボクセルのサイズ
X=Y=Z: 1.68 Å
密度
表面レベル
登録者による: 2.0 / ムービー #1: 2
最小 - 最大
-6.99771738 - 9.615289689999999
平均 (標準偏差)
-0.35500363 (±0.9953196)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
-256
-256
-256
サイズ
512
512
512
Spacing
512
512
512
セル
A=B=C: 860.16 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
1.68
1.68
1.68
M x/y/z
512
512
512
origin x/y/z
0.000
0.000
0.000
length x/y/z
860.160
860.160
860.160
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-147
-147
-146
NX/NY/NZ
294
294
294
MAP C/R/S
1
2
3
start NC/NR/NS
-256
-256
-256
NC/NR/NS
512
512
512
D min/max/mean
-6.998
9.615
-0.355
-
添付データ
-
試料の構成要素
-
全体 : Dengue virus serotype 2 strain New Guinea-C complexed with Fab fr...
全体
名称: Dengue virus serotype 2 strain New Guinea-C complexed with Fab fragments of human antibody 2D22.
要素
試料: Dengue virus serotype 2 strain New Guinea-C complexed with Fab fragments of human antibody 2D22.
ウイルス: Dengue Virus serotype 2 (デング熱ウイルス)
タンパク質・ペプチド: Antigen-binding fragment of human antibody 2D22
-
超分子 #1000: Dengue virus serotype 2 strain New Guinea-C complexed with Fab fr...
超分子
名称: Dengue virus serotype 2 strain New Guinea-C complexed with Fab fragments of human antibody 2D22. タイプ: sample / ID: 1000 / Number unique components: 2
分子 #1: Antigen-binding fragment of human antibody 2D22
分子
名称: Antigen-binding fragment of human antibody 2D22 / タイプ: protein_or_peptide / ID: 1 詳細: 180 copies of Fab 2D22 molecule bind to virus surface コピー数: 180 / 組換発現: No
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: Human / 細胞: memory B-cell
分子量
理論値: 50 KDa
-
実験情報
-
構造解析
手法
クライオ電子顕微鏡法
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: 8 / 詳細: 10 mM Tris-HCl pH 8.0, 120 mM NaCl and 1 mM EDTA
グリッド
詳細: ultra-thin carbon-coated lacey carbon grid
凍結
凍結剤: ETHANE / チャンバー内湿度: 100 % / 装置: FEI VITROBOT MARK IV 手法: blotted with filter paper for 2 seconds prior to snap freezing
-
電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
日付
2014年3月28日
撮影
カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 実像数: 87 / 平均電子線量: 18 e/Å2