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.
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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: 2.17 Å3/Da / 溶媒含有率: 43.28 %
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: 7.3 詳細: 0.2000M CaAcetate, 20.0000% PEG-3350, No Buffer pH 7.3, NANODROP, VAPOR DIFFUSION, SITTING DROP, temperature 277K
モノクロメーター: Double crystal monochromator / プロトコル: SINGLE WAVELENGTH / 単色(M)・ラウエ(L): M / 散乱光タイプ: x-ray
放射波長
波長: 0.97911 Å / 相対比: 1
反射
解像度: 2.11→43.033 Å / Num. obs: 167009 / % possible obs: 99.8 % / Observed criterion σ(I): -3 / 冗長度: 3.8 % / Biso Wilson estimate: 25.198 Å2 / Rmerge(I) obs: 0.122 / Net I/σ(I): 8.25
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
2.11-2.19
0.597
2.1
66321
17620
99.9
2.19-2.27
0.491
2.6
57277
15211
99.9
2.27-2.38
0.4
3.2
66723
17682
99.9
2.38-2.5
0.324
3.8
60374
15935
99.9
2.5-2.66
0.255
4.8
64875
17112
99.9
2.66-2.86
0.194
6.3
61517
16213
99.9
2.86-3.15
0.133
8.8
64080
16877
99.9
3.15-3.6
0.081
13.2
62682
16533
99.7
3.6-4.53
0.056
18.1
63478
16763
99.6
4.53-43.033
0.059
19.4
63570
17041
99.5
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位相決定
位相決定
手法: 単波長異常分散
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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
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: 単波長異常分散 / 解像度: 2.11→43.033 Å / Cor.coef. Fo:Fc: 0.968 / Cor.coef. Fo:Fc free: 0.946 / Occupancy max: 1 / Occupancy min: 0.5 / SU B: 4.695 / SU ML: 0.122 / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.227 / ESU R Free: 0.169 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. DI-METALS (ZINC) AT ACTIVE SITES WERE CONFIRMED WITH X-RAY FLUORESCENE SPECTROSCOPY, ANOMALOUS DIFFERENCE FOURIERS AND COORDINATION GEOMETRY. 4.CALCIUM (CA), ETHYLENE GLYCOL (EDO) AND CHLORIDE (CL) MODELED WERE PRESENT IN CRYSTLLIZATION/CRYO CONDITIONS. 5. DUE TO THE THIN-SHELL SELECTION METHOD, THE HIGHEST SHELL DID NOT CONTAIN ANY FREE REFLECTIONS. THE 1137 REFLECTIONS IN THE SHELL 2.20-2.21 A HAD AN R-FREE OF 0.26.
Rfactor
反射数
%反射
Selection details
Rfree
0.193
8604
5.2 %
THIN SHELLS
Rwork
0.151
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obs
0.153
166973
99.83 %
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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