THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG FOLLOWED BY THE TARGET ...THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG FOLLOWED BY THE TARGET SEQUENCE. DNA SEQUENCING OF THE CLONED CONSTRUCT SHOWS AN ARGININE AT POSITION 244 INSTEAD OF A GLYCINE. THE ARGININE AT POSITION 244 IS SUPPORTED BY THE ELECTRON DENSITY. THE AUTHORS DO NOT KNOW IF THIS GLY TO ARG DIFFERENCE IS DUE TO AN ERROR IN THE DB SEQUENCE, A MUTATION IN THE GENOMIC DNA (SAME SOURCE STRAIN AS USED FOR GENOME SEQUENCING) OR A MUTATION INTRODUCED BY PCR.
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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: 2
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試料調製
結晶
ID
マシュー密度 (Å3/Da)
溶媒含有率 (%)
解説
1
2.71
54.58
THE STRUCTURE WAS SOLVED BY 3-WAVELENGTH MAD METHOD USING 2.4 A DATA FROM ANOTHER CRYSTAL. THESE PHASES WERE USED AS RESTRAINTS IN THE CURRENT REFINEMENT.
解像度: 2.15→29.761 Å / Num. obs: 39885 / % possible obs: 99.9 % / 冗長度: 3.7 % / Biso Wilson estimate: 34.533 Å2 / Rmerge(I) obs: 0.111 / Rsym value: 0.111 / Net I/σ(I): 4.124
反射 シェル
Diffraction-ID: 1,2
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
2.15-2.21
3.8
0.719
1
10975
2923
0.719
100
2.21-2.27
3.7
0.709
0.4
10482
2842
0.709
100
2.27-2.33
3.7
0.532
1.1
10547
2814
0.532
100
2.33-2.4
3.8
0.427
1.7
10124
2691
0.427
100
2.4-2.48
3.8
0.373
1.9
9907
2630
0.373
100
2.48-2.57
3.8
0.306
2.3
9583
2541
0.306
100
2.57-2.67
3.8
0.247
2.5
9283
2469
0.247
100
2.67-2.78
3.8
0.222
1.7
8896
2371
0.222
100
2.78-2.9
3.8
0.166
4.1
8585
2277
0.166
100
2.9-3.04
3.8
0.137
5.1
8064
2139
0.137
100
3.04-3.21
3.8
0.118
5.9
7805
2078
0.118
100
3.21-3.4
3.8
0.101
6.2
7292
1942
0.101
100
3.4-3.63
3.7
0.091
4.3
6853
1843
0.091
100
3.63-3.93
3.7
0.081
5.4
6376
1714
0.081
100
3.93-4.3
3.7
0.065
9.5
5977
1595
0.065
100
4.3-4.81
3.8
0.061
10
5314
1417
0.061
100
4.81-5.55
3.7
0.065
9.5
4736
1270
0.065
100
5.55-6.8
3.7
0.073
8.2
3939
1070
0.073
100
6.8-9.62
3.7
0.064
9.1
3040
832
0.064
99.1
9.62-29.76
3.4
0.059
10
1452
427
0.059
91.3
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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
5.2.0019
精密化
PHENIX
精密化
SHELX
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
XDS
データ削減
XSCALE
データスケーリング
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: 2.15→29.761 Å / Cor.coef. Fo:Fc: 0.96 / Cor.coef. Fo:Fc free: 0.938 / Occupancy max: 1 / Occupancy min: 0.5 / SU B: 11.62 / SU ML: 0.152 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.205 / ESU R Free: 0.177 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 RECORDS CONTAIN RESIDUAL B FACTORS ONLY. 4.NADPH,DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, (NDP) WAS MODELED INTO THE PUTATIVE ACTIVE SITE ON EACH SUBUNIT IN THE ASYMMETRIC UNIT. 5. BICINE (BCN) AND (4R)-2-METHYLPENTANE-2,4-DIOL (MRD) FROM THE CRYSTALLIZATION
Rfactor
反射数
%反射
Selection details
Rfree
0.226
2003
5 %
RANDOM
Rwork
0.183
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obs
0.186
39845
99.84 %
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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