ジャーナル: Nat Struct Mol Biol / 年: 2008 タイトル: Binding of a neutralizing antibody to dengue virus alters the arrangement of surface glycoproteins. 著者: Shee-Mei Lok / Victor Kostyuchenko / Grant E Nybakken / Heather A Holdaway / Anthony J Battisti / Soila Sukupolvi-Petty / Dagmar Sedlak / Daved H Fremont / Paul R Chipman / John T Roehrig / ...著者: Shee-Mei Lok / Victor Kostyuchenko / Grant E Nybakken / Heather A Holdaway / Anthony J Battisti / Soila Sukupolvi-Petty / Dagmar Sedlak / Daved H Fremont / Paul R Chipman / John T Roehrig / Michael S Diamond / Richard J Kuhn / Michael G Rossmann / 要旨: The monoclonal antibody 1A1D-2 has been shown to strongly neutralize dengue virus serotypes 1, 2 and 3, primarily by inhibiting attachment to host cells. A crystal structure of its antigen binding ...The monoclonal antibody 1A1D-2 has been shown to strongly neutralize dengue virus serotypes 1, 2 and 3, primarily by inhibiting attachment to host cells. A crystal structure of its antigen binding fragment (Fab) complexed with domain III of the viral envelope glycoprotein, E, showed that the epitope would be partially occluded in the known structure of the mature dengue virus. Nevertheless, antibody could bind to the virus at 37 degrees C, suggesting that the virus is in dynamic motion making hidden epitopes briefly available. A cryo-electron microscope image reconstruction of the virus:Fab complex showed large changes in the organization of the E protein that exposed the epitopes on two of the three E molecules in each of the 60 icosahedral asymmetric units of the virus. The changes in the structure of the viral surface are presumably responsible for inhibiting attachment to cells.
全体 : Fab Fragment of MAb 1A1D-2 complexed with Dengue 2 virus
全体
名称: Fab Fragment of MAb 1A1D-2 complexed with Dengue 2 virus
要素
試料: Fab Fragment of MAb 1A1D-2 complexed with Dengue 2 virus
ウイルス: Dengue virus 2 (デング熱ウイルス)
タンパク質・ペプチド: Fab fragment of MAb 1A1D-2
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超分子 #1000: Fab Fragment of MAb 1A1D-2 complexed with Dengue 2 virus
超分子
名称: Fab Fragment of MAb 1A1D-2 complexed with Dengue 2 virus タイプ: sample / ID: 1000 集合状態: The Fab molecule binds to two of the three E proteins in the icosahedral assymmetric unit Number unique components: 2
pH: 7.6 / 詳細: 12 mM Tris-HCl, 120 mM NaCl, 1 mM EDTA
グリッド
詳細: 400 mesh copper grid
凍結
凍結剤: ETHANE / 装置: HOMEMADE PLUNGER 詳細: Vitrification instrument: Guillotine-style plunge freezeing device 手法: A small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding a few microliters of the sample is held in place at the bottom of a plunger by the means of fine ...手法: A small vial of ethane is placed inside a larger liquid nitrogen reservoir. The grid holding a few microliters of the sample is held in place at the bottom of a plunger by the means of fine tweezers. Once the ethane in the vial is completely frozen, it needs to be slightly melted. When the liquid ethane is ready, a piece of filter paper is then pressed against the sample to blot of excess buffer, sufficient to leave a thin layer on the grid. After a predetermined time, the filter paper is removed, and the plunger is allowed to drop into the liquid ethane. Once the grid enters the liquid ethane, the sample is rapidly frozen, and the grid is transferred under liquid nitrogen to a storage box immersed liquid nitrogen for later use in the microscope.