ジャーナル: Cell Rep / 年: 2015 タイトル: Neutralizing Monoclonal Antibodies Block Chikungunya Virus Entry and Release by Targeting an Epitope Critical to Viral Pathogenesis. 著者: Jing Jin / Nathan M Liss / Dong-Hua Chen / Maofu Liao / Julie M Fox / Raeann M Shimak / Rachel H Fong / Daniel Chafets / Sonia Bakkour / Sheila Keating / Marina E Fomin / Marcus O Muench / ...著者: Jing Jin / Nathan M Liss / Dong-Hua Chen / Maofu Liao / Julie M Fox / Raeann M Shimak / Rachel H Fong / Daniel Chafets / Sonia Bakkour / Sheila Keating / Marina E Fomin / Marcus O Muench / Michael B Sherman / Benjamin J Doranz / Michael S Diamond / Graham Simmons / 要旨: We evaluated the mechanism by which neutralizing human monoclonal antibodies inhibit chikungunya virus (CHIKV) infection. Potently neutralizing antibodies (NAbs) blocked infection at multiple steps ...We evaluated the mechanism by which neutralizing human monoclonal antibodies inhibit chikungunya virus (CHIKV) infection. Potently neutralizing antibodies (NAbs) blocked infection at multiple steps of the virus life cycle, including entry and release. Cryo-electron microscopy structures of Fab fragments of two human NAbs and chikungunya virus-like particles showed a binding footprint that spanned independent domains on neighboring E2 subunits within one viral spike, suggesting a mechanism for inhibiting low-pH-dependent membrane fusion. Detailed epitope mapping identified amino acid E2-W64 as a critical interaction residue. An escape mutation (E2-W64G) at this residue rendered CHIKV attenuated in mice. Consistent with these data, CHIKV-E2-W64G failed to emerge in vivo under the selection pressure of one of the NAbs, IM-CKV063. As our study suggests that antibodies engaging the residue E2-W64 can potently inhibit CHIKV at multiple stages of infection, antibody-based therapies or immunogens that target this region might have protective value.
ダウンロード / ファイル: emd_6457.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 CHIKV VLP in complex with Fab fragment of neutralizing antibody IM-CKV063
ボクセルのサイズ
X=Y=Z: 1.93 Å
密度
表面レベル
登録者による: 0.012 / ムービー #1: 0.012
最小 - 最大
-0.01421068 - 0.0305851
平均 (標準偏差)
0.00083652 (±0.00567338)
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
512
512
512
Spacing
512
512
512
セル
A=B=C: 988.16 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
1.93
1.93
1.93
M x/y/z
512
512
512
origin x/y/z
0.000
0.000
0.000
length x/y/z
988.160
988.160
988.160
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-190
-190
-189
NX/NY/NZ
380
380
380
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
512
512
512
D min/max/mean
-0.014
0.031
0.001
-
添付データ
-
試料の構成要素
-
全体 : Chikungunya (S27) virus-like particle in complex with Fab fragmen...
全体
名称: Chikungunya (S27) virus-like particle in complex with Fab fragment of neutralizing antibody IM-CKV063
要素
試料: Chikungunya (S27) virus-like particle in complex with Fab fragment of neutralizing antibody IM-CKV063
ウイルス: Chikungunya virus (チクングニヤウイルス)
タンパク質・ペプチド: IM-CKV063 Fab
-
超分子 #1000: Chikungunya (S27) virus-like particle in complex with Fab fragmen...
超分子
名称: Chikungunya (S27) virus-like particle in complex with Fab fragment of neutralizing antibody IM-CKV063 タイプ: sample / ID: 1000 / Number unique components: 2
名称: IM-CKV063 Fab / タイプ: protein_or_peptide / ID: 1 / 組換発現: No / データベース: NCBI
由来(天然)
生物種: Mus musculus (ハツカネズミ) / 別称: mouse
-
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
濃度
2 mg/mL
緩衝液
pH: 7.4 / 詳細: 20 mM HEPES, 150 mM NaCl
グリッド
詳細: Quantifoil copper grids (R2/1, 200 mesh)
凍結
凍結剤: ETHANE / チャンバー内湿度: 90 % / チャンバー内温度: 120 K / 装置: FEI VITROBOT MARK III / 手法: Blot for 2.5 seconds before plunging.
-
電子顕微鏡法
顕微鏡
FEI TECNAI F20
温度
最低: 96 K / 最高: 98 K / 平均: 97 K
アライメント法
Legacy - 非点収差: Objective lens astigmatism was corrected at 29,000 times magnification.
日付
2015年6月16日
撮影
カテゴリ: CCD / フィルム・検出器のモデル: GATAN K2 (4k x 4k) / デジタル化 - サンプリング間隔: 5.0 µm / 実像数: 96 / 平均電子線量: 18 e/Å2 詳細: Every image is the sum of 30 frames (200 ms/frame) recorded by the K2 Summit direct electron detector after the drift correction and damage compensation based on the whole frame. ビット/ピクセル: 32