THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH ...THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH TEV PROTEASE LEAVING ONLY A GLYCINE (0) FOLLOWED BY THE TARGET SEQUENCE. THE DNA SEQUENCING OF THE CLONED CONSTRUCT SHOWS A LEUCINE AT POSITION 127 INSTEAD OF A VALINE. THE LEUCINE AT POSITION 127 IS SUPPORTED BY THE ELECTRON DENSITY.
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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: 2.29 Å3/Da / 溶媒含有率: 46.25 %
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: 7 詳細: 31.0% 2-methyl-2,4-pentanediol, 0.2M sodium chloride, 0.1M TRIS pH 7.0, NANODROP, VAPOR DIFFUSION, SITTING DROP, temperature 277K
モノクロメーター: Single crystal Si(111) bent monochromator (ho rizontal 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
0.97968
1
2
0.97917
1
3
0.91837
1
反射
解像度: 2.5→29.074 Å / Num. obs: 15426 / % possible obs: 93.6 % / Observed criterion σ(I): -3 / Biso Wilson estimate: 49.009 Å2 / Rmerge(I) obs: 0.047 / Net I/σ(I): 11.12
反射 シェル
解像度 (Å)
最高解像度 (Å)
Rmerge(I) obs
Mean I/σ(I) obs
Num. measured obs
Num. unique obs
% possible all
2.5-2.59
0.352
2.3
3928
2486
81.3
2.59-2.69
0.298
2.8
4363
2653
88.6
2.69-2.81
0.212
3.9
4633
2739
91.1
2.81-2.96
0.156
5.2
5084
2897
92.5
2.96-3.15
0.118
6.7
5392
2959
95.1
3.15-3.39
0.071
10.3
5569
2949
96.2
3.39-3.73
0.047
14.2
5738
2989
97.2
3.73-4.26
0.033
18.5
5766
2981
98
4.26-5.35
0.029
21.1
5855
3032
98.4
5.35
0.027
22.5
5933
3064
97.2
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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
5.2.0019
精密化
PHENIX
精密化
SOLVE
位相決定
MolProbity
3beta29
モデル構築
XSCALE
データスケーリング
PDB_EXTRACT
3.004
データ抽出
XDS
データ削減
精密化
構造決定の手法: 多波長異常分散 / 解像度: 2.5→29.074 Å / Cor.coef. Fo:Fc: 0.921 / Cor.coef. Fo:Fc free: 0.898 / SU B: 11.675 / SU ML: 0.26 / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.691 / ESU R Free: 0.324 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.
Rfactor
反射数
%反射
Selection details
Rfree
0.277
772
5 %
RANDOM
Rwork
0.241
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-
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obs
0.243
15414
97.85 %
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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