- PDB-1z1v: NMR structure of the Saccharomyces cerevisiae Ste50 SAM domain -
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基本情報
登録情報
データベース: PDB / ID: 1z1v
タイトル
NMR structure of the Saccharomyces cerevisiae Ste50 SAM domain
要素
STE50 protein
キーワード
CELL CYCLE / all helix protein / SAM domain
機能・相同性
機能・相同性情報
osmosensory signaling pathway via Sho1 osmosensor / signal transduction involved in filamentous growth / SAM domain binding / pheromone-dependent signal transduction involved in conjugation with cellular fusion / protein kinase regulator activity / establishment of cell polarity / p38MAPK cascade / regulation of cell cycle / cytoplasm 類似検索 - 分子機能
分子量: 9309.657 Da / 分子数: 1 / 断片: ste50 SAM domain (residues 32-107) / 由来タイプ: 組換発現 由来: (組換発現) Saccharomyces cerevisiae (パン酵母) 解説: this protein fragment was cloned into the NdeI and BamHI sites of pET15b creating an aminoterminal hexahistidine tagged protein with an intervening thrombin protease cleavage site. After ...解説: this protein fragment was cloned into the NdeI and BamHI sites of pET15b creating an aminoterminal hexahistidine tagged protein with an intervening thrombin protease cleavage site. After thrombin treatment, the non-native residues GSH remained amino terminal to the native Ste50 sequence 遺伝子: STE50 / プラスミド: pET15b / 生物種 (発現宿主): Escherichia coli / 発現宿主: Escherichia coli BL21(DE3) (大腸菌) / 株 (発現宿主): BL21(DE3) / 参照: UniProt: P25344
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実験情報
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実験
実験
手法: 溶液NMR
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
3D 13C-separated NOESY
1
2
1
3D 15N-separated NOESY
NMR実験の詳細
Text: This structure was determined using standard 3D heteronuclear techniques
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試料調製
詳細
内容: 0.8 mM Ste50 SAM U-15N,13C, 20 mM sodium phosphate buffer, pH 7.8, 150 mM sodium chloride, 0.02 % sodium azide, 90% H2O, 10% D2O 溶媒系: 90% H2O/10% D2O
試料状態
イオン強度: 150 mM NaCl / pH: 7.8 / 圧: 1 atm / 温度: 293 K
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NMR測定
放射
プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M
放射波長
相対比: 1
NMRスペクトロメーター
タイプ
製造業者
モデル
磁場強度 (MHz)
Spectrometer-ID
Bruker AVANCE
Bruker
AVANCE
600
1
Varian INOVA
Varian
INOVA
800
2
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解析
NMR software
名称
バージョン
開発者
分類
NMRPipe
Delagio
解析
NMRView
5.2
Johnson
データ解析
CYANA
2
Guntert
構造決定
XPLOR-NIH
2.9.9
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1 詳細: Experimental observations: 615 intermolecular NOE distance restraints, 328 short range NOE distance restraints, 163 medium range NOE distance restraints, 149 long range NOE restraints, 42 ...詳細: Experimental observations: 615 intermolecular NOE distance restraints, 328 short range NOE distance restraints, 163 medium range NOE distance restraints, 149 long range NOE restraints, 42 pairs of hydrogen bond distance restraints and 69 pairs of phi/psi dihedral angle restraints. An initial ensemble of 500 structures were calculated with CYANA 2.0. The top 25 structures with minimum restraint violations were refined in water using XPLOR-NIH and a protocol by C. Spronk. All 25 water refined structures had no NOE violations > 0.5 A and no dihedral violations > 5 degrees. For residues 35-100, the backbone RMSD of the ensemble is 0.75 +/- 0.14 A. Residues 28-32 and 101-107 are disordered.