ジャーナル: Cell / 年: 2008 タイトル: Adenovirus serotype 5 hexon mediates liver gene transfer. 著者: Simon N Waddington / John H McVey / David Bhella / Alan L Parker / Kristeen Barker / Hideko Atoda / Rebecca Pink / Suzanne M K Buckley / Jenny A Greig / Laura Denby / Jerome Custers / Takashi ...著者: Simon N Waddington / John H McVey / David Bhella / Alan L Parker / Kristeen Barker / Hideko Atoda / Rebecca Pink / Suzanne M K Buckley / Jenny A Greig / Laura Denby / Jerome Custers / Takashi Morita / Ivo M B Francischetti / Robson Q Monteiro / Dan H Barouch / Nico van Rooijen / Claudio Napoli / Menzo J E Havenga / Stuart A Nicklin / Andrew H Baker / 要旨: Adenoviruses are used extensively as gene transfer agents, both experimentally and clinically. However, targeting of liver cells by adenoviruses compromises their potential efficacy. In cell culture, ...Adenoviruses are used extensively as gene transfer agents, both experimentally and clinically. However, targeting of liver cells by adenoviruses compromises their potential efficacy. In cell culture, the adenovirus serotype 5 fiber protein engages the coxsackievirus and adenovirus receptor (CAR) to bind cells. Paradoxically, following intravascular delivery, CAR is not used for liver transduction, implicating alternate pathways. Recently, we demonstrated that coagulation factor (F)X directly binds adenovirus leading to liver infection. Here, we show that FX binds to the Ad5 hexon, not fiber, via an interaction between the FX Gla domain and hypervariable regions of the hexon surface. Binding occurs in multiple human adenovirus serotypes. Liver infection by the FX-Ad5 complex is mediated through a heparin-binding exosite in the FX serine protease domain. This study reveals an unanticipated function for hexon in mediating liver gene transfer in vivo.
名称: Factor X / タイプ: ligand / ID: 1 / Name.synonym: FX 詳細: Commercially purified Factor X from Haematologic Technologies Inc. コピー数: 240 / 集合状態: monomer / 組換発現: No
由来(天然)
組織: blood/liver
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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
緩衝液
pH: 7.4 / 詳細: 10 nM Tris 100 mM NaCl 5 mM CaCl
グリッド
詳細: 400 mesh copper R2/2 quantifoils
凍結
凍結剤: ETHANE 手法: 5 ul of virus-ligand complex was loaded onto a freshly glow-discharged grid and blotted until the filter paper detached.
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電子顕微鏡法
顕微鏡
JEOL 1200EXII
温度
平均: 108 K
アライメント法
Legacy - 非点収差: Astigmatism corrected at 100,000 times magnification
詳細
Defocus Pairs were recorded
日付
2007年6月1日
撮影
カテゴリ: FILM / フィルム・検出器のモデル: KODAK SO-163 FILM / デジタル化 - スキャナー: OTHER / デジタル化 - サンプリング間隔: 6.35 µm / 実像数: 21 / 平均電子線量: 10 e/Å2 / 詳細: Images were binned by a factor of 3 / ビット/ピクセル: 16