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 RESIDUES 19-460 OF THE TARGET SEQUENCE
-
実験情報
-
実験
実験
手法: X線回折 / 使用した結晶の数: 1
-
試料調製
結晶
マシュー密度: 2.55 Å3/Da / 溶媒含有率: 51.72 %
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: 8.2 詳細: 0.200M sodium citrate, 20.00% PEG-3350, No Buffer pH 8.2, 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
0.97828
1
3
0.97916
1
反射
解像度: 1.6→28.072 Å / Num. obs: 133836 / % possible obs: 99.7 % / 冗長度: 3 % / Biso Wilson estimate: 16.908 Å2 / Rmerge(I) obs: 0.074 / Rsym value: 0.074 / Net I/σ(I): 6.547
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
1.6-1.64
2.9
0.365
2
28854
9900
0.365
100
1.64-1.69
2.9
0.32
2.3
28185
9649
0.32
100
1.69-1.74
2.9
0.27
2.7
27582
9414
0.27
100
1.74-1.79
2.9
0.217
3.3
26587
9078
0.217
100
1.79-1.85
2.9
0.187
3.8
25916
8823
0.187
100
1.85-1.91
2.9
0.164
4.3
25200
8576
0.164
100
1.91-1.98
2.9
0.14
5
24274
8250
0.14
100
1.98-2.07
2.9
0.114
6
23308
7913
0.114
100
2.07-2.16
3
0.095
7.1
22570
7638
0.095
100
2.16-2.26
3
0.085
7.7
21532
7267
0.085
99.9
2.26-2.39
3
0.081
7.9
20626
6958
0.081
99.9
2.39-2.53
3
0.081
7.4
19336
6515
0.081
99.8
2.53-2.7
3
0.078
7.9
18292
6141
0.078
99.7
2.7-2.92
3
0.067
9.1
17115
5735
0.067
99.6
2.92-3.2
3
0.057
10.4
15815
5266
0.057
99.4
3.2-3.58
3
0.049
12
14359
4757
0.049
99.2
3.58-4.13
3
0.047
12.7
12619
4174
0.047
98.9
4.13-5.06
3
0.052
11.6
10759
3561
0.052
98.7
5.06-7.16
3
0.057
10.9
8218
2731
0.057
98.2
7.16-28.08
3
0.045
14.2
4414
1490
0.045
95.7
-
位相決定
位相決定
手法: 多波長異常分散
-
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
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: 1.6→28.072 Å / Cor.coef. Fo:Fc: 0.975 / Cor.coef. Fo:Fc free: 0.964 / Occupancy max: 1 / Occupancy min: 0.25 / SU B: 2.578 / SU ML: 0.047 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.068 / ESU R Free: 0.071 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 RECORDS CONTAIN 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 RECORDS CONTAIN 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 FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 4. ELECTRON DENSITY INDICATES THAT THE PEPTIDE BOND BETWEEN TYR 226 AND HIS 227 ON BOTH SUBUNITS IN THE ASYMMETRIC UNIT IS IN THE CIS-CONFIGURATION. TYR 226 AND HIS 227 ARE IN THE VICINITY OF THE PUTATIVE ACTIVE SITE. 5. AN UNKNOWN LIGAND (UNL) WAS MODELED INTO THE PUTATIVE ACTIVE IN CHAIN B. 6. 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID (EPE - HEPES) FROM THE PROTEIN BUFFER WAS MODELED IN THE PUTATIVE ACTIVE SITE OF CHAIN A. 7. ETHYLENE GLYCOLS (EDO) USED AS A CRYOPROTECTANT AND SODIUM IONS FROM THE CRYSTALLIZATION SOLUTION WERE MODELED INTO THE STRUCTURE. 8. ELECTRON DENSITY BETWEEN RESUDES 260-266 ON THE B-SUBUNIT WAS DISORDERED, AND THESE RESIDUES WERE NOT MODELED.
Rfactor
反射数
%反射
Selection details
Rfree
0.168
6726
5 %
RANDOM
Rwork
0.139
-
-
-
obs
0.14
133835
99.65 %
-
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