ジャーナル: Proc Natl Acad Sci U S A / 年: 2013 タイトル: Cross-neutralizing human anti-poliovirus antibodies bind the recognition site for cellular receptor. 著者: Zhaochun Chen / Elizabeth R Fischer / Diana Kouiavskaia / Bryan T Hansen / Steven J Ludtke / Bella Bidzhieva / Michelle Makiya / Liane Agulto / Robert H Purcell / Konstantin Chumakov / 要旨: Most structural information about poliovirus interaction with neutralizing antibodies was obtained in the 1980s in studies of mouse monoclonal antibodies. Recently we have isolated a number of ...Most structural information about poliovirus interaction with neutralizing antibodies was obtained in the 1980s in studies of mouse monoclonal antibodies. Recently we have isolated a number of human/chimpanzee anti-poliovirus antibodies and demonstrated that one of them, MAb A12, could neutralize polioviruses of both serotypes 1 and 2. This communication presents data on isolation of an additional cross-neutralizing antibody (F12) and identification of a previously unknown epitope on the surface of poliovirus virions. Epitope mapping was performed by sequencing of antibody-resistant mutants and by cryo-EM of complexes of virions with Fab fragments. The results have demonstrated that both cross-neutralizing antibodies bind the site located at the bottom of the canyon surrounding the fivefold axis of symmetry that was previously shown to interact with cellular poliovirus receptor CD155. However, the same antibody binds to serotypes 1 and 2 through different specific interactions. It was also shown to interact with type 3 poliovirus, albeit with about 10-fold lower affinity, insufficient for effective neutralization. Antibody interaction with the binding site of the cellular receptor may explain its broad reactivity and suggest that further screening or antibody engineering could lead to a universal antibody capable of neutralizing all three serotypes of poliovirus.
A: 430.19998 Å / B: 426.59998 Å / C: 430.19998 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
1.8
1.8
1.8
M x/y/z
239
237
239
origin x/y/z
0.000
0.000
0.000
length x/y/z
430.200
426.600
430.200
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-132
-122
-147
NX/NY/NZ
250
274
261
MAP C/R/S
1
2
3
start NC/NR/NS
41
42
41
NC/NR/NS
239
237
239
D min/max/mean
0.000
2.154
0.252
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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 : Fab Fragment of A12 antibody bound to Type 2 poliovirus
全体
名称: Fab Fragment of A12 antibody bound to Type 2 poliovirus
要素
試料: Fab Fragment of A12 antibody bound to Type 2 poliovirus
ウイルス: Human poliovirus 2 (ポリオウイルス)
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超分子 #1000: Fab Fragment of A12 antibody bound to Type 2 poliovirus
超分子
名称: Fab Fragment of A12 antibody bound to Type 2 poliovirus タイプ: sample / ID: 1000 / Number unique components: 2
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超分子 #1: Human poliovirus 2
超分子
名称: Human poliovirus 2 / タイプ: virus / ID: 1 / NCBI-ID: 12083 / 生物種: Human poliovirus 2 / データベース: NCBI / ウイルスタイプ: VIRION / ウイルス・単離状態: SPECIES / ウイルス・エンベロープ: Yes / ウイルス・中空状態: No
宿主
生物種: Homo sapiens (ヒト) / 別称: VERTEBRATES
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実験情報
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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
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試料調製
濃度
1 mg/mL
グリッド
詳細: 4 uL aliquots of sample were applied to freshly glow-discharged 200 mesh r2/2 Quantifoil copper grids suspended by forceps in the FEI Mark IV Vitrobot.
凍結
凍結剤: ETHANE / チャンバー内湿度: 80 % / チャンバー内温度: 65 K / 装置: FEI VITROBOT MARK IV 手法: Specimens were vitrified, after blotting for 2 seconds with a blot force of 5 at 80% relative humidity, by plunge freezing into liquid ethane.
Particles with a defocus range of 1-5 um were boxed with EMAN2, and then processed using the standard single particle reconstruction procedure with full CTF correction.
CTF補正
詳細: each image
最終 再構成
解像度のタイプ: BY AUTHOR / 解像度: 20.0 Å / 解像度の算出法: FSC 0.5 CUT-OFF / ソフトウェア - 名称: EMAN2 / 使用した粒子像数: 1500