ジャーナル: Proc Natl Acad Sci U S A / 年: 2014 タイトル: Structures of protective antibodies reveal sites of vulnerability on Ebola virus. 著者: Charles D Murin / Marnie L Fusco / Zachary A Bornholdt / Xiangguo Qiu / Gene G Olinger / Larry Zeitlin / Gary P Kobinger / Andrew B Ward / Erica Ollmann Saphire / 要旨: Ebola virus (EBOV) and related filoviruses cause severe hemorrhagic fever, with up to 90% lethality, and no treatments are approved for human use. Multiple recent outbreaks of EBOV and the likelihood ...Ebola virus (EBOV) and related filoviruses cause severe hemorrhagic fever, with up to 90% lethality, and no treatments are approved for human use. Multiple recent outbreaks of EBOV and the likelihood of future human exposure highlight the need for pre- and postexposure treatments. Monoclonal antibody (mAb) cocktails are particularly attractive candidates due to their proven postexposure efficacy in nonhuman primate models of EBOV infection. Two candidate cocktails, MB-003 and ZMAb, have been extensively evaluated in both in vitro and in vivo studies. Recently, these two therapeutics have been combined into a new cocktail named ZMapp, which showed increased efficacy and has been given compassionately to some human patients. Epitope information and mechanism of action are currently unknown for most of the component mAbs. Here we provide single-particle EM reconstructions of every mAb in the ZMapp cocktail, as well as additional antibodies from MB-003 and ZMAb. Our results illuminate key and recurring sites of vulnerability on the EBOV glycoprotein and provide a structural rationale for the efficacy of ZMapp. Interestingly, two of its components recognize overlapping epitopes and compete with each other for binding. Going forward, this work now provides a basis for strategic selection of next-generation antibody cocktails against Ebola and related viruses and a model for predicting the impact of ZMapp on potential escape mutations in ongoing or future Ebola outbreaks.
ダウンロード / ファイル: emd_6152.map.gz / 形式: CCP4 / 大きさ: 1.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Reconstruction of Ebola virus glycoprotein bound to Fab fragments of c13C6 and c4G7
ボクセルのサイズ
X=Y=Z: 4.1 Å
密度
表面レベル
登録者による: 3.65 / ムービー #1: 3.65
最小 - 最大
-6.99864483 - 11.645090100000001
平均 (標準偏差)
0.00309111 (±0.99991232)
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
80
80
80
Spacing
80
80
80
セル
A=B=C: 328.0 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
4.1
4.1
4.1
M x/y/z
80
80
80
origin x/y/z
0.000
0.000
0.000
length x/y/z
328.000
328.000
328.000
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
0
0
0
NX/NY/NZ
96
96
80
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
80
80
80
D min/max/mean
-6.999
11.645
0.003
-
添付データ
-
試料の構成要素
-
全体 : Fab fragment of c13C6 and c4G7 chimerized human monoclonal IgG1 a...
全体
名称: Fab fragment of c13C6 and c4G7 chimerized human monoclonal IgG1 antibody bound to Ebola virus glycoprotein
要素
試料: Fab fragment of c13C6 and c4G7 chimerized human monoclonal IgG1 antibody bound to Ebola virus glycoprotein
タンパク質・ペプチド: Ebola virus glycoprotein
タンパク質・ペプチド: chimerized human IgG1 antigen binding fragment c13C6
タンパク質・ペプチド: chimerized human IgG1 antigen binding fragment c4G7
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超分子 #1000: Fab fragment of c13C6 and c4G7 chimerized human monoclonal IgG1 a...
超分子
名称: Fab fragment of c13C6 and c4G7 chimerized human monoclonal IgG1 antibody bound to Ebola virus glycoprotein タイプ: sample / ID: 1000 集合状態: Six Fab fragments bound to a single, trimeric GP (two Fabs per protomer) Number unique components: 3
分子 #2: chimerized human IgG1 antigen binding fragment c13C6
分子
名称: chimerized human IgG1 antigen binding fragment c13C6 タイプ: protein_or_peptide / ID: 2 / Name.synonym: c13C6 Fab / コピー数: 3 / 集合状態: monomer / 組換発現: Yes
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: human
組換発現
生物種: Nicotiana benthamiana (ナス科)
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分子 #3: chimerized human IgG1 antigen binding fragment c4G7
分子
名称: chimerized human IgG1 antigen binding fragment c4G7 / タイプ: protein_or_peptide / ID: 3 / Name.synonym: c4G7 Fab / コピー数: 3 / 集合状態: monomer / 組換発現: Yes
由来(天然)
生物種: Homo sapiens (ヒト) / 別称: human
組換発現
生物種: Nicotiana benthamiana (ナス科)
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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
0.03 mg/mL
緩衝液
pH: 7.4 / 詳細: 20 mM Tris, 150 mM NaCl
染色
タイプ: NEGATIVE 詳細: To prepare negative stain grids, a 4 uL aliquot of each complex, which had been diluted to a concentration of ~0.03 ug/mL with TBS buffer, was placed for 15 seconds onto carbon-coated 400 Cu ...詳細: To prepare negative stain grids, a 4 uL aliquot of each complex, which had been diluted to a concentration of ~0.03 ug/mL with TBS buffer, was placed for 15 seconds onto carbon-coated 400 Cu mesh grids that had been plasma cleaned for 20 s (Gatan), blotted off on the edge of the grid, then immediately stained for 30 s with 4 uL of 2% uranyl formate. The stain was blotted off on the edge of the grid and the grid was allowed to dry.