- EMDB-3334: Cryo electron microscopy of a complex of Tor-323RFP and Lst8 -
+
データを開く
IDまたはキーワード:
読み込み中...
-
基本情報
登録情報
データベース: EMDB / ID: EMD-3334
タイトル
Cryo electron microscopy of a complex of Tor-323RFP and Lst8
マップデータ
Reconstruction of a complex of variant Tor-323RFP and Lst8.
試料
試料: Complex of Tor-323RFP with Lst8
タンパク質・ペプチド: Target of rapamycin (Tor)
タンパク質・ペプチド: Lst8
タンパク質・ペプチド: Red fluorescent protein (RFP)
キーワード
cryo-EM / Tor / Lst8 / mTOR / kinase / PIKK / S/T protein kinase / TORC1 / mTORC1 / RFP
機能・相同性
TORC1 complex / Green fluorescent protein, GFP / Green fluorescent protein-related / Green fluorescent protein / Green fluorescent protein / bioluminescence / generation of precursor metabolites and energy / Red fluorescent protein drFP583
ジャーナル: Nat Commun / 年: 2016 タイトル: Tor forms a dimer through an N-terminal helical solenoid with a complex topology. 著者: Domagoj Baretić / Alex Berndt / Yohei Ohashi / Christopher M Johnson / Roger L Williams / 要旨: The target of rapamycin (Tor) is a Ser/Thr protein kinase that regulates a range of anabolic and catabolic processes. Tor is present in two complexes, TORC1 and TORC2, in which the Tor-Lst8 ...The target of rapamycin (Tor) is a Ser/Thr protein kinase that regulates a range of anabolic and catabolic processes. Tor is present in two complexes, TORC1 and TORC2, in which the Tor-Lst8 heterodimer forms a common sub-complex. We have determined the cryo-electron microscopy (EM) structure of Tor bound to Lst8. Two Tor-Lst8 heterodimers assemble further into a dyad-symmetry dimer mediated by Tor-Tor interactions. The first 1,300 residues of Tor form a HEAT repeat-containing α-solenoid with four distinct segments: a highly curved 800-residue N-terminal 'spiral', followed by a 400-residue low-curvature 'bridge' and an extended 'railing' running along the bridge leading to the 'cap' that links to FAT region. This complex topology was verified by domain insertions and offers a new interpretation of the mTORC1 structure. The spiral of one TOR interacts with the bridge of another, which together form a joint platform for the Regulatory Associated Protein of TOR (RAPTOR) regulatory subunit.
名称: Red fluorescent protein (RFP) / タイプ: protein_or_peptide / ID: 3 / Name.synonym: RFP, dsRed, FP583 詳細: The tandem RFP was inserted between residues G323 and G324 of K. marxianus Target of rapamycin (Tor) polypeptide chain. コピー数: 4 / 集合状態: Dimer / 組換発現: Yes
由来(天然)
生物種: Discosoma sp. (イソギンチャク) / 別称: Coral anemones
分子量
理論値: 54 KDa
組換発現
生物種: Kluyveromyces marxianus (酵母)
配列
UniProtKB: Red fluorescent protein drFP583
-
実験情報
-
構造解析
手法
クライオ電子顕微鏡法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
濃度
2.5 mg/mL
緩衝液
pH: 7.4 詳細: 50 mM Hepes pH 7.4 (23 deg C), 75 mM KCl, 250 mM NaCl, 0.3 % (v/v) CHAPS, 1 mM TCEP
グリッド
詳細: Quantifoil Au R 0.6/1.0 or Au R 1.2/1.3, 300 mesh grids, blotted for 11-13 s at 4 deg C
凍結
凍結剤: ETHANE / 装置: OTHER / 手法: 11-13 s at 4 deg C
-
電子顕微鏡法
顕微鏡
FEI POLARA 300
日付
2015年9月22日
撮影
カテゴリ: CCD フィルム・検出器のモデル: FEI FALCON II (4k x 4k) 実像数: 678 / 平均電子線量: 40 e/Å2
Processed with RELION. CTF corrected each particle.
CTF補正
詳細: Each particle (Gctf)
最終 再構成
想定した対称性 - 点群: C1 (非対称) / 解像度のタイプ: BY AUTHOR / 解像度: 9.1 Å / 解像度の算出法: OTHER / ソフトウェア - 名称: RELION 詳細: Final map was calculated after masked classification with density subtraction on a single tandem RFP tag. 使用した粒子像数: 10386