ジャーナル: PLoS Pathog / 年: 2017 タイトル: Two classes of protective antibodies against Pseudorabies virus variant glycoprotein B: Implications for vaccine design. 著者: Xiangdong Li / Fanli Yang / Xule Hu / Feifei Tan / Jianxun Qi / Ruchao Peng / Min Wang / Yan Chai / Liying Hao / Junhua Deng / Chenyu Bai / Juan Wang / Hao Song / Shuguang Tan / Guangwen Lu / ...著者: Xiangdong Li / Fanli Yang / Xule Hu / Feifei Tan / Jianxun Qi / Ruchao Peng / Min Wang / Yan Chai / Liying Hao / Junhua Deng / Chenyu Bai / Juan Wang / Hao Song / Shuguang Tan / Guangwen Lu / George F Gao / Yi Shi / Kegong Tian / 要旨: Pseudorabies virus (PRV) belongs to the Herpesviridae family, and is an important veterinary pathogen. Highly pathogenic PRV variants have caused severe epidemics in China since 2011, causing huge ...Pseudorabies virus (PRV) belongs to the Herpesviridae family, and is an important veterinary pathogen. Highly pathogenic PRV variants have caused severe epidemics in China since 2011, causing huge economic losses. To tackle the epidemics, we identified a panel of mouse monoclonal antibodies (mAbs) against PRV glycoprotein B (gB) that effectively block PRV infection. Among these 15 mAbs, fourteen of them block PRV entry in a complement-dependent manner. The remaining one, 1H1 mAb, however can directly neutralize the virus independent of complement and displays broad-spectrum neutralizing activities. We further determined the crystal structure of PRV gB and mapped the epitopes of these antibodies on the structure. Interestingly, all the complement-dependent neutralizing antibodies bind gB at the crown region (domain IV). In contrast, the epitope of 1H1 mAb is located at the bottom of domain I, which includes the fusion loops, indicating 1H1 mAb might neutralize the virus by interfering with the membrane fusion process. Our studies demonstrate that gB contains multiple B-cell epitopes in its crown and base regions and that antibodies targeting different epitopes block virus infection through different mechanisms. These findings would provide important clues for antiviral drug design and vaccine development.
ダウンロード / ファイル: emd_6841.map.gz / 形式: CCP4 / 大きさ: 125 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
EM density map
ボクセルのサイズ
X=Y=Z: 1.36 Å
密度
表面レベル
登録者による: 0.0046 / ムービー #1: 0.0046
最小 - 最大
-0.009441859 - 0.036231082
平均 (標準偏差)
0.000062737985 (±0.0014707975)
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
320
320
320
Spacing
320
320
320
セル
A=B=C: 435.2 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
1.36
1.36
1.36
M x/y/z
320
320
320
origin x/y/z
0.000
0.000
0.000
length x/y/z
435.200
435.200
435.200
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
320
320
320
D min/max/mean
-0.009
0.036
0.000
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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 : Pseudorabies virus glycoprotein B in complex with Fab fragments f...
全体
名称: Pseudorabies virus glycoprotein B in complex with Fab fragments from a neutralizing antibody
要素
複合体: Pseudorabies virus glycoprotein B in complex with Fab fragments from a neutralizing antibody
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超分子 #1: Pseudorabies virus glycoprotein B in complex with Fab fragments f...
超分子
名称: Pseudorabies virus glycoprotein B in complex with Fab fragments from a neutralizing antibody タイプ: complex / ID: 1 / 親要素: 0 詳細: The Fab fragment was generated by proteolytic cleavage of IgG antibody. The Pseudorabies virus glycoprotein B was expressed using the baculovirus expression system. The two proteins were ...詳細: The Fab fragment was generated by proteolytic cleavage of IgG antibody. The Pseudorabies virus glycoprotein B was expressed using the baculovirus expression system. The two proteins were purified separately and mixed to constitute complex samples in solution.
由来(天然)
生物種: Suid herpesvirus 1 (ヘルペスウイルス)
組換発現
生物種: Spodoptera frugiperda (ツマジロクサヨトウ) 組換細胞: Sf9
分子量
実験値: 300 KDa
-
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
濃度
0.02 mg/mL
緩衝液
pH: 8 / 詳細: 20 mM Tris-HCl, pH 8.0 150 mM NaCl
染色
タイプ: NEGATIVE / 材質: Uranyl acetate 詳細: The specimen was stained by 1% Uranyl acetate for 1 min and air-dried before data acquisition.