THE CONSTRUCT (RESIDUES 18-506) WAS EXPRESSED WITH AN N-TERMINAL PURIFICATION TAG ...THE CONSTRUCT (RESIDUES 18-506) 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. DNA SEQUENCING OF THE CLONED CONSTRUCT REVEALS A GLUTAMATE AT POSITION 418 INSTEAD OF AN ALANINE. THE GLUTAMATE AT POSITION 418 IS SUPPORTED BY 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: 2.09 Å3/Da / 溶媒含有率: 41.09 % 解説: DATA WERE SCALED USING XSCALE WITH FRIEDEL PAIRS KEPT AS SEPARATE WHEN COMPUTING R-MERGE, COMPLETENESS AND
結晶化
温度: 293 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 magnesium chloride, 30.0000% polyethylene glycol 4000, 0.1M TRIS pH 8.5, Additive: 0.001 M glucose, NANODROP', VAPOR DIFFUSION, SITTING DROP, temperature 293K
解像度: 2→29.182 Å / Num. obs: 61624 / % possible obs: 95.4 % / Observed criterion σ(I): -3 / Biso Wilson estimate: 15.487 Å2 / Rmerge(I) obs: 0.096 / Net I/σ(I): 6.35
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
2-2.07
0.35
1.9
17313
11175
92.1
2.07-2.15
0.28
2.4
17679
11336
94.5
2.15-2.25
0.23
2.9
18707
12017
94.8
2.25-2.37
0.192
3.5
18547
11927
95.3
2.37-2.52
0.159
4.2
18524
11951
95.5
2.52-2.71
0.131
5
17974
11608
96.1
2.71-2.99
0.097
6.3
18933
12241
96.4
2.99-3.42
0.065
8.8
18400
11925
96.9
3.42-4.3
0.04
13.5
18190
11842
96.6
4.3-29.182
0.035
14.5
18185
11903
95.3
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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
5.5.0109
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
SHELXD
位相決定
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: 2→29.182 Å / Cor.coef. Fo:Fc: 0.953 / Cor.coef. Fo:Fc free: 0.922 / Occupancy max: 1 / Occupancy min: 0.22 / SU B: 7.669 / SU ML: 0.112 / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R Free: 0.17 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. HEPES (EPE, 4-(2-HYDROXYETHYL)-1-PIPERAZINEETHANESULFONIC ACID) AND CHLORIDE (CL) MODELED ARE PRESENT PROTEIN BUFFER.
Rfactor
反射数
%反射
Selection details
Rfree
0.216
3125
5.1 %
RANDOM
Rwork
0.164
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obs
0.167
61624
98.23 %
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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: MASK