SEQUENCE THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS ... SEQUENCE 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. RESIDUES GLU 161 AND ASP 163 WERE MUTATED TO ALA IN THE EXPRESSED CONSTRUCT. THE PROTEIN WAS REDUCTIVELY METHYLATED PRIOR TO CRYSTALLIZATION.
モノクロメーター: Single crystal Si(111) bent (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
0.97913
1
3
0.97883
1
反射
解像度: 1.9→29.514 Å / Num. obs: 104039 / % possible obs: 99.8 % / Observed criterion σ(I): -3 / Biso Wilson estimate: 27.76 Å2 / Rmerge(I) obs: 0.088 / Net I/σ(I): 8.27
反射 シェル
解像度 (Å)
Rmerge(I) obs
Mean I/σ(I) obs
Num. measured obs
Diffraction-ID
% possible all
1.9-1.97
0.614
1.7
58613
1
99
1.97-2.05
0.475
2.2
58348
1
100
2.05-2.14
0.374
2.8
55978
1
99.9
2.14-2.25
0.313
3.4
56921
1
100
2.25-2.39
0.254
4.2
58046
1
100
2.39-2.58
0.208
5.1
60565
1
100
2.58-2.84
0.152
7
58722
1
100
2.84-3.25
0.089
11.6
59002
1
100
3.25-4.08
0.049
19.4
58330
1
100
4.08-29.514
0.036
25.1
59512
1
99.4
-
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
XSCALE
データスケーリング
PDB_EXTRACT
3
データ抽出
MAR345
CCD
データ収集
XDS
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: 1.9→29.514 Å / Cor.coef. Fo:Fc: 0.968 / Cor.coef. Fo:Fc free: 0.96 / SU B: 3.495 / SU ML: 0.055 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.079 / ESU R Free: 0.079 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 RECORD CONTAINS 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 RECORD CONTAINS 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 TO ACCOUNT FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 4. FOUR NI IONS, SEVEN CHLORIDE IONS, FOUR SULFATE IONS AND TWO GLYCEROL MOLECULES WERE MODELED. THE PRESENCE OF THE NI ATOMS ARE SUPPORTED BY X-RAY FLUORESCENCE MEASUREMENTS, ANOMALOUS DIFFERENCE FOURIERS AND GEOMETRY. 5. RESIDUES 1 AND 265 TO 269 IN EACH SUBUNIT ARE DISORDERED AND NOT MODELED IN THE STRUCTURE. 6. THE DISUFIDE BOND BETWEEN CYS 212 FROM EACH SUBUNIT IS PARTIALLY REDUCED.
Rfactor
反射数
%反射
Selection details
Rfree
0.181
5190
5 %
RANDOM
Rwork
0.16
-
-
-
obs
0.161
103941
99.92 %
-
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