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基本情報
登録情報 | データベース: EMDB / ID: EMD-5927 | |||||||||
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タイトル | Kinetic and Structural Analysis of Coxsackievirus B3 Receptor Interactions and Formation of the A-particle | |||||||||
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![]() | Coxsackievirus B3 complexed with CAR: virus ![]() | |||||||||
![]() | coxsackievirus b3 / cvb3 / ![]() ![]() ![]() | |||||||||
機能・相同性 | ![]() AV node cell-bundle of His cell adhesion involved in cell communication / cell adhesive protein binding involved in AV node cell-bundle of His cell communication / AV node cell to bundle of His cell communication / homotypic cell-cell adhesion / epithelial structure maintenance / regulation of AV node cell action potential / gamma-delta T cell activation / apicolateral plasma membrane / germ cell migration / transepithelial transport ...AV node cell-bundle of His cell adhesion involved in cell communication / cell adhesive protein binding involved in AV node cell-bundle of His cell communication / AV node cell to bundle of His cell communication / homotypic cell-cell adhesion / epithelial structure maintenance / regulation of AV node cell action potential / gamma-delta T cell activation / apicolateral plasma membrane / germ cell migration / transepithelial transport / cardiac muscle fiber development / cell-cell junction organization / negative regulation of cardiac muscle cell proliferation / ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() ![]() | |||||||||
生物種 | ![]() ![]() ![]() ![]() ![]() | |||||||||
手法 | ![]() ![]() | |||||||||
![]() | Organtini LJ / Makhov AM / Conway JF / Hafenstein S / Carson SD | |||||||||
![]() | ![]() タイトル: Kinetic and structural analysis of coxsackievirus B3 receptor interactions and formation of the A-particle. 著者: Lindsey J Organtini / Alexander M Makhov / James F Conway / Susan Hafenstein / Steven D Carson / ![]() 要旨: The coxsackievirus and adenovirus receptor (CAR) has been identified as the cellular receptor for group B coxsackieviruses, including serotype 3 (CVB3). CAR mediates infection by binding to CVB3 and ...The coxsackievirus and adenovirus receptor (CAR) has been identified as the cellular receptor for group B coxsackieviruses, including serotype 3 (CVB3). CAR mediates infection by binding to CVB3 and catalyzing conformational changes in the virus that result in formation of the altered, noninfectious A-particle. Kinetic analyses show that the apparent first-order rate constant for the inactivation of CVB3 by soluble CAR (sCAR) at physiological temperatures varies nonlinearly with sCAR concentration. Cryo-electron microscopy (cryo-EM) reconstruction of the CVB3-CAR complex resulted in a 9.0-Å resolution map that was interpreted with the four available crystal structures of CAR, providing a consensus footprint for the receptor binding site. The analysis of the cryo-EM structure identifies important virus-receptor interactions that are conserved across picornavirus species. These conserved interactions map to variable antigenic sites or structurally conserved regions, suggesting a combination of evolutionary mechanisms for receptor site preservation. The CAR-catalyzed A-particle structure was solved to a 6.6-Å resolution and shows significant rearrangement of internal features and symmetric interactions with the RNA genome. Importance: This report presents new information about receptor use by picornaviruses and highlights the importance of attaining at least an ∼9-Å resolution for the interpretation of cryo-EM ...Importance: This report presents new information about receptor use by picornaviruses and highlights the importance of attaining at least an ∼9-Å resolution for the interpretation of cryo-EM complex maps. The analysis of receptor binding elucidates two complementary mechanisms for preservation of the low-affinity (initial) interaction of the receptor and defines the kinetics of receptor-catalyzed conformational change to the A-particle. | |||||||||
構造検証レポート | ![]() ![]() ![]() ![]() | |||||||||
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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: 1.25 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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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
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試料の構成要素
-全体 Coxsackievirus B3 complexed with CAR
全体 | 名称: Coxsackievirus B3 complexed with CAR / 詳細: purified virus and receptor complex in solution / 構成要素数: 2 / オリゴマーの状態: icosahedral virus |
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分子量 | 実験値: 7 MDa |
-構成要素 #1: ウイルス, Human coxsackievirus B3
ウイルス | 名称: Human coxsackievirus B3 / 別称: CVB3 / クラス: VIRION / 詳細: Virus was incubated with excess CAR at 4 degrees C. / エンベロープを持つか: No / 中空か: No / 単離: STRAIN |
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分子量 | 実験値: 7 MDa |
生物種 | 生物種: ![]() ![]() ![]() 株: CVB3/28 |
由来(天然) | 宿主: ![]() ![]() |
殻 #1 | 要素名: VP1-4 / 直径: 300 Å / T番号(三角分割数): 1 |
-構成要素 #2: タンパク質, Coxsackievirus and adenovirus receptor
タンパク質 | 名称: Coxsackievirus and adenovirus receptor / 別称: CAR![]() |
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由来 | 生物種: ![]() ![]() |
由来(合成) | 発現系: ![]() ![]() ![]() |
外部リンク | UniProt: Coxsackievirus and adenovirus receptor |
-実験情報
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試料調製
試料 | 試料の状態: 粒子 / 手法: ![]() |
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試料溶液 | 試料濃度: 0.1 mg/mL / 緩衝液: 50 mM MES, 100 mM NaCl / pH: 6 |
支持膜 | glow-discharged holey carbon Quantifoil electron microscopy grids |
凍結 | 装置: FEI VITROBOT MARK III / 凍結剤: ETHANE-PROPANE MIXTURE / 温度: 95 K / 湿度: 95 % |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Tecnai F20 / 画像提供: FEI Company |
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![]() | 顕微鏡: FEI TECNAI F20 / 日付: 2012年8月1日 |
電子銃 | 電子線源: FIELD EMISSION GUN![]() |
レンズ | 倍率: 50000 X (nominal), 50000 X (calibrated) / 非点収差: CTFFIND3 / Cs: 2 mm / 撮影モード: BRIGHT FIELD![]() |
試料ホルダ | モデル: GATAN LIQUID NITROGEN |
カメラ | 検出器: KODAK SO-163 FILM |
-画像取得
画像取得 | デジタル画像の数: 96 / Scanner: NIKON SUPER COOLSCAN 9000 / サンプリングサイズ: 6.35 µm |
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画像解析
![]() | 手法: ![]() ![]() |
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3次元再構成 | アルゴリズム: Common Lines / ソフトウェア: EMAN, AUTO3DEM / CTF補正: AUTO3DEM / 解像度: 9 Å / 解像度の算出法: FSC 0.5, semi-independent |
-原子モデル構築
モデリング #1 | ソフトウェア: Situs / 精密化のプロトコル: rigid body / 当てはまり具合の基準: correlation coefficient / 精密化に使用した空間: REAL 詳細: The four available CAR structures were fit into the receptor density separately. 利用したPDBモデル: 1F5W 鎖ID: B |
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モデリング #2 | ソフトウェア: Situs / 精密化のプロトコル: rigid body / 当てはまり具合の基準: correlation coefficient / 精密化に使用した空間: REAL 詳細: The four available CAR structures were fit into the receptor density separately. 利用したPDBモデル: 1KAC 鎖ID: B |
モデリング #3 | ソフトウェア: Situs / 精密化のプロトコル: rigid body / 当てはまり具合の基準: correlation coefficient / 精密化に使用した空間: REAL 詳細: The four available CAR structures were fit into the receptor density separately. 利用したPDBモデル: 3JZ7 鎖ID: K |
モデリング #4 | ソフトウェア: Situs / 精密化のプロトコル: rigid body / 当てはまり具合の基準: correlation coefficient / 精密化に使用した空間: REAL 詳細: The four available CAR structures were fit into the receptor density separately. 利用したPDBモデル: 3MJ7 鎖ID: S |
得られたモデル |