CRYSTAL PACKING ANALYSIS AND ANALYTICAL SIZE EXCLUSION CHROMATOGRAPHY COUPLED WITH STATIC LIGHT SCATTERING SUPPORT THE ASSIGNMENT OF A TRIMER AS A SIGNIFICANT OLIGOMERIC FORM IN SOLUTION.
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要素
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タンパク質 , 1種, 1分子 A
#1: タンパク質
Putativearomatic-ringhydroxylatingdioxygenase / Probable Ring hydroxylating alpha subunit
THIS CONSTRUCT WAS EXPRESSED WITH AN N-TERMINAL PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS ...THIS CONSTRUCT WAS EXPRESSED WITH AN N-TERMINAL PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH TEV PROTEASE LEAVING ONLY A GLYCINE (0) FOLLOWED BY THE TARGET SEQUENCE.
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実験情報
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実験
実験
手法: X線回折 / 使用した結晶の数: 1
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試料調製
結晶
マシュー密度: 2.85 Å3/Da / 溶媒含有率: 56.85 %
結晶化
温度: 277 K / 手法: 蒸気拡散法, シッティングドロップ法 / pH: 8.29 詳細: 23.8000% polyethylene glycol monomethyl ether 2000, 0.0100M nickel (II) chloride, 0.1M TRIS pH 8.29, NANODROP', VAPOR DIFFUSION, SITTING DROP, temperature 277K
モノクロメーター: Single crystal Si(111) bent monochromator (horizontal focusing) プロトコル: MAD / 単色(M)・ラウエ(L): M / 散乱光タイプ: x-ray
放射波長
ID
波長 (Å)
相対比
1
0.91837
1
2
0.97949
1
3
0.97895
1
反射
解像度: 1.8→28.155 Å / Num. obs: 48265 / % possible obs: 99.3 % / Observed criterion σ(I): -3 / 冗長度: 6.31 % / Biso Wilson estimate: 22.737 Å2 / Rmerge(I) obs: 0.06 / Net I/σ(I): 14.33
反射 シェル
解像度 (Å)
Rmerge(I) obs
Mean I/σ(I) obs
Num. measured obs
Num. unique obs
% possible all
1.8-1.86
0.577
2.16
28099
8783
98.9
1.86-1.94
0.421
3
32555
10184
99.1
1.94-2.03
0.274
4.4
30765
9585
99.2
2.03-2.13
0.199
6.1
28346
8822
99.2
2.13-2.27
0.14
8.4
32011
9942
99.3
2.27-2.44
0.1
11.4
29634
9189
99.4
2.44-2.69
0.076
14.8
31247
9675
99.5
2.69-3.07
0.052
20.7
29995
9290
99.6
3.07-3.87
0.034
31.4
30874
9574
99.7
3.87-28.155
0.024
40.4
30919
9582
99.5
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位相決定
位相決定
手法: 多波長異常分散
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解析
ソフトウェア
名称
バージョン
分類
NB
REFMAC
5.6.0073
精密化
PHENIX
精密化
SHELX
位相決定
MolProbity
3beta29
モデル構築
XSCALE
データスケーリング
PDB_EXTRACT
3.006
データ抽出
XDS
データ削減
autoSHARP
位相決定
精密化
構造決定の手法: 多波長異常分散 / 解像度: 1.8→28.155 Å / Cor.coef. Fo:Fc: 0.965 / Cor.coef. Fo:Fc free: 0.951 / Occupancy max: 1 / Occupancy min: 0.3 / SU B: 3.589 / SU ML: 0.063 / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R Free: 0.098 立体化学のターゲット値: MAXIMUM LIKELIHOOD WITH PHASES 詳細: 1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. A MET-INHIBITION PROTOCOL WAS USED FOR SELENOMETHIONINE INCORPORATION DURING PROTEIN EXPRESSION. THE OCCUPANCY OF THE SE ATOMS IN THE ...詳細: 1. HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2. A MET-INHIBITION PROTOCOL WAS USED FOR SELENOMETHIONINE INCORPORATION DURING PROTEIN EXPRESSION. THE OCCUPANCY OF THE SE ATOMS IN THE MSE RESIDUES WAS REDUCED TO 0.75 FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 3. ATOM RECORD CONTAINS SUM OF TLS AND RESIDUAL B FACTORS. ANISOU RECORD CONTAINS SUM OF TLS AND RESIDUAL U FACTORS. 4. NICKEL (NI), CHLORIDE (CL) AND TROMETHAMINE (TRS; TRIS) FROM THE CRYSTALLIZATION SOLUTION AND GLYCEROL (GOL) FROM THE CRYOPROTECTANT SOLUTION HAVE BEEN MODELED. 5. AN IRON-SULPHUR [2FE-2S] CLUSTER HAS BEEN MODELED. 6. THE PRESENCE OF IRON AND NICKEL HAVE BEEN VERIFIED BY X-RAY FLUORESCENCE AND BY CALCULATING ANOMALOUS DIFFERENCE FOURIER MAPS FROM DATA COLLECTED BELOW AND ABOVE THE IRON AND NICKEL K-SHELL ABSORPTION EDGES.
Rfactor
反射数
%反射
Selection details
Rfree
0.186
2418
5 %
RANDOM
Rwork
0.154
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obs
0.156
48240
99.98 %
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溶媒の処理
イオンプローブ半径: 0.8 Å / 減衰半径: 0.8 Å / VDWプローブ半径: 1.2 Å / 溶媒モデル: BABINET MODEL WITH MASK