+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-4013 | |||||||||
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タイトル | Structure of a hexagonal assembly of the PFV glycoprotein from the iFuse Env mutant by cryo-electron microscopy | |||||||||
マップデータ | None | |||||||||
試料 |
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生物種 | Human spumaretrovirus (ウイルス) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 8.8 Å | |||||||||
データ登録者 | Effantin G | |||||||||
引用 | ジャーナル: PLoS Pathog / 年: 2016 タイトル: Cryo-electron Microscopy Structure of the Native Prototype Foamy Virus Glycoprotein and Virus Architecture. 著者: Grégory Effantin / Leandro F Estrozi / Nick Aschman / Patricia Renesto / Nicole Stanke / Dirk Lindemann / Guy Schoehn / Winfried Weissenhorn / 要旨: Foamy viruses (FV) belong to the genus Spumavirus, which forms a distinct lineage in the Retroviridae family. Although the infection in natural hosts and zoonotic transmission to humans is ...Foamy viruses (FV) belong to the genus Spumavirus, which forms a distinct lineage in the Retroviridae family. Although the infection in natural hosts and zoonotic transmission to humans is asymptomatic, FVs can replicate well in human cells making it an attractive gene therapy vector candidate. Here we present cryo-electron microscopy and (cryo-)electron tomography ultrastructural data on purified prototype FV (PFV) and PFV infected cells. Mature PFV particles have a distinct morphology with a capsid of constant dimension as well as a less ordered shell of density between the capsid and the membrane likely formed by the Gag N-terminal domain and the cytoplasmic part of the Env leader peptide gp18LP. The viral membrane contains trimeric Env glycoproteins partly arranged in interlocked hexagonal assemblies. In situ 3D reconstruction by subtomogram averaging of wild type Env and of a Env gp48TM- gp80SU cleavage site mutant showed a similar spike architecture as well as stabilization of the hexagonal lattice by clear connections between lower densities of neighboring trimers. Cryo-EM was employed to obtain a 9 Å resolution map of the glycoprotein in its pre-fusion state, which revealed extensive trimer interactions by the receptor binding subunit gp80SU at the top of the spike and three central helices derived from the fusion protein subunit gp48TM. The lower part of Env, presumably composed of interlaced parts of gp48TM, gp80SU and gp18LP anchors the spike at the membrane. We propose that the gp48TM density continues into three central transmembrane helices, which interact with three outer transmembrane helices derived from gp18LP. Our ultrastructural data and 9 Å resolution glycoprotein structure provide important new insights into the molecular architecture of PFV and its distinct evolutionary relationship with other members of the Retroviridae. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_4013.map.gz | 9 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-4013-v30.xml emd-4013.xml | 9.8 KB 9.8 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_4013.png | 104.1 KB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-4013 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-4013 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_4013_validation.pdf.gz | 223.4 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_4013_full_validation.pdf.gz | 222.5 KB | 表示 | |
XML形式データ | emd_4013_validation.xml.gz | 5.2 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4013 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-4013 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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-マップ
ファイル | ダウンロード / ファイル: emd_4013.map.gz / 形式: CCP4 / 大きさ: 9.6 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | None | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.94 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
-試料の構成要素
-全体 : Human spumaretrovirus
全体 | 名称: Human spumaretrovirus (ウイルス) |
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要素 |
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-超分子 #1: Human spumaretrovirus
超分子 | 名称: Human spumaretrovirus / タイプ: virus / ID: 1 / 親要素: 0 / 詳細: Mutant in the Env polyprotein / NCBI-ID: 11963 / 生物種: Human spumaretrovirus / 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 |
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Host system | 生物種: Homo sapiens (ヒト) / 組換プラスミド: pcoPG4, pcoPE32, pcoPP |
分子量 | 理論値: 330 KDa |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
緩衝液 | pH: 7.5 |
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凍結 | 凍結剤: ETHANE |
詳細 | The iFuse mutant is a variant of Env where the furine cleavage site between the SUrface (gp80 SU ) and TransMembrane (gp48 TM ) domains of Env has been mutated. This results in a partially processed glycoprotein as the cleavage between the LP and SU domains is preserved. Particles are released at nearly wild type level from cells but are non-infectious. |
-電子顕微鏡法
顕微鏡 | FEI TECNAI F30 |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 平均電子線量: 30.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
実験機器 | モデル: Tecnai F30 / 画像提供: FEI Company |