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 CLONED CONSTRUCT CONTAINS RESIDUES 27-312 OF THE FULL LENGTH PROTEIN.
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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: 2.47 Å3/Da / 溶媒含有率: 50.14 %
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: 8.5 詳細: 0.2000M MgCl2, 20.0000% PEG-8000, 0.1M TRIS pH 8.5, 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
0.9791
1
3
0.97858
1
反射
解像度: 1.68→29.374 Å / Num. obs: 71903 / % possible obs: 99.6 % / 冗長度: 3.7 % / Biso Wilson estimate: 11.88 Å2 / Rmerge(I) obs: 0.132 / Rsym value: 0.132 / Net I/σ(I): 8.6
反射 シェル
Diffraction-ID: 1
解像度 (Å)
冗長度 (%)
Rmerge(I) obs
Mean I/σ(I) obs
Num. measured all
Num. unique all
Rsym value
% possible all
1.68-1.72
3.7
0.541
1.4
19446
5273
0.541
100
1.72-1.77
3.7
0.47
1.6
18869
5129
0.47
100
1.77-1.82
3.7
0.404
1.9
18445
4992
0.404
100
1.82-1.88
3.7
0.341
2.2
17938
4857
0.341
100
1.88-1.94
3.7
0.302
2.5
17502
4737
0.302
100
1.94-2.01
3.7
0.24
3
16879
4562
0.24
100
2.01-2.08
3.7
0.206
3.6
16416
4432
0.206
99.9
2.08-2.17
3.7
0.176
4.2
15754
4256
0.176
99.9
2.17-2.27
3.7
0.155
4.7
15099
4076
0.155
99.9
2.27-2.38
3.7
0.144
5
14579
3910
0.144
99.8
2.38-2.5
3.7
0.13
5.5
13769
3718
0.13
99.8
2.5-2.66
3.7
0.123
5.6
13142
3534
0.123
99.7
2.66-2.84
3.7
0.117
5.8
12232
3311
0.117
99.5
2.84-3.07
3.7
0.102
6.3
11408
3090
0.102
99.4
3.07-3.36
3.7
0.084
7.5
10515
2848
0.084
99.2
3.36-3.76
3.7
0.07
9
9488
2570
0.07
98.9
3.76-4.34
3.7
0.063
9.6
8442
2287
0.063
98.6
4.34-5.31
3.7
0.065
9.2
7182
1946
0.065
98.4
5.31-7.51
3.6
0.073
8.7
5528
1521
0.073
97.5
7.51-29.37
3.4
0.063
10
2943
854
0.063
94.6
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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
5.5.0102
精密化
PHENIX
精密化
SOLVE
位相決定
MolProbity
3beta29
モデル構築
SCALA
3.2.5
データスケーリング
PDB_EXTRACT
3.006
データ抽出
MOSFLM
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: 1.68→29.374 Å / Cor.coef. Fo:Fc: 0.962 / Cor.coef. Fo:Fc free: 0.946 / Occupancy max: 1 / Occupancy min: 0.25 / SU B: 3.519 / SU ML: 0.053 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.084 / ESU R Free: 0.085 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.ATOM RECORD CONTAINS RESIDUAL B FACTORS ONLY. 3.A MET-INHIBITION PROTOCOL WAS USED FOR SELENOMETHIONINE INCORPORATION DURING PROTEIN ...詳細: 1.HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS. 2.ATOM RECORD CONTAINS RESIDUAL B FACTORS ONLY. 3.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 TO ACCOUNT FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 4.MAGNESIUM ATOMS (MG) AND POLYETHYLENE GLYCOL (PEG) FROM THE CRYSTALLIZATION SOLUTION ARE MODELED INTO THE STRUCTURE. 5. TLS PARAMETERS WERE ASSIGNED WITH THE AID OF THE TLS MOTION DETERMINATION SERVER. 6. A RAMACHANDRAN OUTLIER (A106) IS SUPPORTED BY CLEARLY DEFINED DENSITY.
Rfactor
反射数
%反射
Selection details
Rfree
0.182
3624
5 %
RANDOM
Rwork
0.15
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obs
0.152
71861
99.41 %
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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: 1.4 Å / 溶媒モデル: MASK