Hsc70-induced Changes in Clathrin-Auxilin Cage Structure Suggest a Role for Clathrin Light Chains in Cage Disassembly
マップデータ
Map of Clathrin-Auxilin complex. Sharpened to match a Fourier amplitude profile derived from a model of the hexagonal barrel (the biological unit) created from the crystal structure atomic model pdb id1XI5. Fourier filtered to 30 Angstroms.
ジャーナル: J Mol Biol / 年: 2004 タイトル: Location of auxilin within a clathrin cage. 著者: Corinne J Smith / Timothy R Dafforn / Helen Kent / Catherine A Sims / Kavita Khubchandani-Aswani / Lin Zhang / Helen R Saibil / Barbara M F Pearse / 要旨: The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism ...The Dna J homologue, auxilin, acts as a co-chaperone for Hsc70 in the uncoating of clathrin-coated vesicles during endocytosis. Biochemical studies have aided understanding of the uncoating mechanism but until now there was no structural information on how auxilin interacts with the clathrin cage. Here we have determined the three-dimensional structure of a complex of auxilin with clathrin cages by cryo-electron microscopy and single particle analysis. We show that auxilin forms a discrete shell of density on the inside of the clathrin cage. Peptide competition assays confirm that a candidate clathrin box motif in auxilin, LLGLE, can bind to a clathrin construct containing the beta-propeller domain and also displace the well-characterised LLNLD clathrin box motif derived from the beta-adaptin hinge region. The means by which auxilin could both aid clathrin coat assembly and displace clathrin from AP2 during uncoating is discussed.
ダウンロード / ファイル: emd_2411.map.gz / 形式: CCP4 / 大きさ: 62.5 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
Map of Clathrin-Auxilin complex. Sharpened to match a Fourier amplitude profile derived from a model of the hexagonal barrel (the biological unit) created from the crystal structure atomic model pdb id1XI5. Fourier filtered to 30 Angstroms.
pH: 6 詳細: 20 mM Mes (pH 6.0), 2 mM magnesium acetate, 25 mM KCl, 10 mM (NH4)2SO4, 1 mM DTT
凍結
凍結剤: ETHANE / 装置: HOMEMADE PLUNGER
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電子顕微鏡法 #1
Microscopy ID
1
顕微鏡
FEI/PHILIPS CM200FEG
日付
2000年2月2日
撮影
デジタル化 - スキャナー: ZEISS SCAI
電子線
加速電圧: 200 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
試料ステージ
試料ホルダーモデル: GATAN LIQUID NITROGEN
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電子顕微鏡法 #2
Microscopy ID
2
顕微鏡
FEI/PHILIPS CM200FEG
日付
2000年2月23日
撮影
デジタル化 - スキャナー: ZEISS SCAI
電子線
加速電圧: 200 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
試料ステージ
試料ホルダーモデル: GATAN LIQUID NITROGEN
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画像解析
CTF補正
詳細: Whole micrographs
最終 再構成
想定した対称性 - 点群: D6 (2回x6回 2面回転対称) アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 31.0 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: MRC, SPIDER, FREALIGN 詳細: Sharpened to match a Fourier amplitude profile derived from a model of the hexagonal barrel (the biological unit) created from the crystal structure atomic model pdb id1XI5. Fourier filtered to 30 Angstroms. 使用した粒子像数: 1745