ジャーナル: Nat Struct Mol Biol / 年: 2019 タイトル: Cryo-EM structure of the ClpXP protein degradation machinery. 著者: Christos Gatsogiannis / Dora Balogh / Felipe Merino / Stephan A Sieber / Stefan Raunser / 要旨: The ClpXP machinery is a two-component protease complex that performs targeted protein degradation in bacteria and mitochondria. The complex consists of the AAA+ chaperone ClpX and the peptidase ClpP. ...The ClpXP machinery is a two-component protease complex that performs targeted protein degradation in bacteria and mitochondria. The complex consists of the AAA+ chaperone ClpX and the peptidase ClpP. The hexameric ClpX utilizes the energy of ATP binding and hydrolysis to engage, unfold and translocate substrates into the catalytic chamber of tetradecameric ClpP, where they are degraded. Formation of the complex involves a symmetry mismatch, because hexameric AAA+ rings bind axially to the opposing stacked heptameric rings of the tetradecameric ClpP. Here we present the cryo-EM structure of ClpXP from Listeria monocytogenes. We unravel the heptamer-hexamer binding interface and provide novel insight into the ClpX-ClpP cross-talk and activation mechanism. Comparison with available crystal structures of ClpP and ClpX in different states allows us to understand important aspects of the complex mode of action of ClpXP and provides a structural framework for future pharmacological applications.
名称: LmClpXP1/2 / タイプ: complex / ID: 2 / 親要素: 1 / 含まれる分子: all 詳細: LmCLP1 and LmCLP2 assemble into two heptameric rings which stack face to face to form the LmClpP1/2 heterocomplex. The chaperone ClpX forms a hexamer, that associates with the LmClpP2 heptamer.
凍結剤: ETHANE / チャンバー内湿度: 90 % / 装置: GATAN CRYOPLUNGE 3 詳細: the sample was blotted after 45sec of incubation time..
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電子顕微鏡法
顕微鏡
FEI TITAN KRIOS
撮影
フィルム・検出器のモデル: FEI FALCON III (4k x 4k) 検出モード: INTEGRATING / 撮影したグリッド数: 1 / 実像数: 3200 / 平均露光時間: 2.0 sec. / 平均電子線量: 114.0 e/Å2 詳細: Images were collected in movie-mode at a frame rate of 50msec
電子線
加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN
電子光学系
倍率(補正後): 112807 / 照射モード: OTHER / 撮影モード: BRIGHT FIELD / Cs: 0.0 mm / 倍率(公称値): 59000