ジャーナル: Structure / 年: 2008 タイトル: Structure of a copper pump suggests a regulatory role for its metal-binding domain. 著者: Chen-Chou Wu / William J Rice / David L Stokes / 要旨: P-type ATPases play an important role in Cu homeostasis, which provides sufficient Cu for metalloenzyme biosynthesis but prevents oxidative damage of free Cu to the cell. The P(IB) group of P-type ...P-type ATPases play an important role in Cu homeostasis, which provides sufficient Cu for metalloenzyme biosynthesis but prevents oxidative damage of free Cu to the cell. The P(IB) group of P-type ATPases includes ATP-dependent pumps of Cu and other transition metal ions, and it is distinguished from other family members by the presence of N-terminal metal-binding domains (MBD). We have determined structures of two constructs of a Cu pump from Archaeoglobus fulgidus (CopA) by cryoelectron microscopy of tubular crystals, which reveal the overall architecture and domain organization of the molecule. By comparing these structures, we localized its N-terminal MBD within the cytoplasmic domains that use ATP hydrolysis to drive the transport cycle. We have built a pseudoatomic model by fitting existing crystallographic structures into the cryoelectron microscopy maps for CopA, which suggest a Cu-dependent regulatory role for the MBD.
pH: 6.1 詳細: 50 mM MES, 25 mM Na2SO4, 25 mM K2SO4, 10 mM MgSO4, 2 mM 2-mercaptoethanol, 200 uM BCDS,
グリッド
詳細: 300 mesh holey carbon
凍結
凍結剤: ETHANE / チャンバー内湿度: 75 % / チャンバー内温度: 100 K / 装置: HOMEMADE PLUNGER / 詳細: Vitrification instrument: home made plunger 手法: Blot for 2-5 seconds before plunging. Plunge in cold room.
詳細
crystals grown at 45 to 55 C
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電子顕微鏡法
顕微鏡
FEI/PHILIPS CM200FEG/UT
温度
平均: 100 K
アライメント法
Legacy - 非点収差: objective astigmatism corrected at 200,000X mag