1. THIS CONSTRUCT WAS EXPRESSED WITH AN N-TERMINAL PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG ...1. THIS CONSTRUCT 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. 2. THE PROTEIN WAS REDUCTIVELY METHYLATED PRIOR TO CRYSTALLIZATION.
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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: 2.61 Å3/Da / 溶媒含有率: 52.8 % 解説: THE STATISTICS REPORTED IN REMARK 200 WERE COMPUTED WITH XSCALE WITH FRIEDEL PAIRS KEPT SEPARATE.
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: 6 詳細: 5.0000% polyethylene glycol 3000, 40.0000% polyethylene glycol 400, 0.1M MES pH 6.0, 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
0.97871
1
3
0.97831
1
Reflection twin
Crystal-ID
ID
Operator
Domain-ID
Fraction
1
1
H, K, L
1
0.772
1
1
K, H, -L
2
0.228
反射
解像度: 1.49→26.832 Å / Num. obs: 26406 / % possible obs: 96.1 % / Observed criterion σ(I): -3 / Biso Wilson estimate: 17.255 Å2 / Rmerge(I) obs: 0.03 / Net I/σ(I): 13.48
反射 シェル
解像度 (Å)
Rmerge(I) obs
Mean I/σ(I) obs
Num. measured obs
Num. unique obs
Diffraction-ID
% possible all
1.49-1.54
0.315
2
6919
4712
1
94.9
1.54-1.6
0.242
2.7
7317
4986
1
95.3
1.6-1.68
0.177
3.6
8071
5506
1
95.6
1.68-1.77
0.121
5.2
7582
5156
1
96.1
1.77-1.88
0.08
7.6
7372
5020
1
96.1
1.88-2.02
0.054
11.2
7174
4886
1
96.3
2.02-2.23
0.036
16.9
7771
5301
1
96.7
2.23-2.55
0.028
22.2
7357
5029
1
96.1
2.55-3.21
0.022
28
7399
5059
1
96.7
3.21-26.832
0.017
34.9
7513
5154
1
97.4
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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
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: 1.49→26.832 Å / Cor.coef. Fo:Fc: 0.981 / Cor.coef. Fo:Fc free: 0.973 / Occupancy max: 1 / Occupancy min: 0.15 / SU B: 1.693 / SU ML: 0.027 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.009 / ESU R Free: 0.01 / 立体化学のターゲット値: MAXIMUM LIKELIHOOD 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. THE DATA IS TWINNED. THE FINAL TWIN FRACTION WAS REFINED TO 0.228 USING THE OPERATOR "K,H,-L". 5.THE PROTEIN WAS SUBJECTED TO REDUCTIVE METHYLATION PRIOR TO CRYSTALLIZATION. THE LYSINE RESIDUES HAVE BEEN MODELED AS N-DIMETHYL-LYSINE (MLY). 6. CYSTEINE 93 IS OXIDIZED AND IS MODELED AS S-OXY CYSTEINE (CSX). 7. CHLORIDE ION (CL) IS MODELED FROM THE PROTEIN BUFFER SOLUTION.
Rfactor
反射数
%反射
Selection details
Rfree
0.14
1355
5.1 %
RANDOM
Rwork
0.122
-
-
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obs
0.123
26404
99.81 %
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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: BABINET MODEL WITH MASK