ジャーナル: To be published タイトル: Crystal structure of uncharacterized protein (JCVI_PEP_1096685590403) from an environmental metagenome (unidentified marine microbe), Sorcerer II Global Ocean Sampling experiment at 2.53 A resolution 著者: Joint Center for Structural Genomics (JCSG)
BIOMOLECULE: 1 THIS ENTRY CONTAINS THE CRYSTALLOGRAPHIC ASYMMETRIC UNIT WHICH CONSISTS OF 5 ... BIOMOLECULE: 1 THIS ENTRY CONTAINS THE CRYSTALLOGRAPHIC ASYMMETRIC UNIT WHICH CONSISTS OF 5 CHAIN(S). SEE REMARK 350 FOR INFORMATION ON GENERATING THE BIOLOGICAL MOLECULE(S). SIZE EXCLUSION CHROMATOGRAPHY WITH STATIC LIGHT SCATTERING SUPPORTS THE ASSIGNMENT OF A PENTAMER AS A SIGNIFICANT OLIGOMERIZATION STATE IN SOLUTION.
Remark 999
SEQUENCE 1. THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS ... SEQUENCE 1. THE CONSTRUCT WAS EXPRESSED WITH A PURIFICATION TAG MGSDKIHHHHHHENLYFQG. THE TAG WAS REMOVED WITH TEV PROTEASE LEAVING ONLY A GLYCINE, FOLLOWED BY THE TARGET SEQUENCE. 2. THE SEQUENCE OF THE PROTEIN WAS NOT AVAILABLE IN THE UNIPROT DATABASE AT THE TIME OF DEPOSITION. 3. THE PRODUCT OF THE EXPRESSED SYNTHETIC GENE WAS BASED ON THE PREDICTED SEQUENCE OF ACCESSION ID JCVI_PEP_1096685590403 FROM THE J. CRAIG VENTER INSTITUTE.
SIZE EXCLUSION CHROMATOGRAPHY WITH STATIC LIGHT SCATTERING SUPPORTS THE ASSIGNMENT OF A PENTAMER AS A SIGNIFICANT OLIGOMERIZATION STATE IN SOLUTION.
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要素
#1: タンパク質
uncharacterizedprotein
分子量: 23422.406 Da / 分子数: 5 / 由来タイプ: 組換発現 由来: (組換発現) uncultured marine organism (環境試料) 遺伝子: SYNTHETIC GENE: The gene product was based on JCVI_PEP_1096685590403 from the Sorcerer II Global Ocean Sampling experiment プラスミド: speedET / 発現宿主: Escherichia coli (大腸菌) / 株 (発現宿主): HK100
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
0.97949
1
反射
解像度: 2.53→29.921 Å / Num. obs: 39917 / % possible obs: 99.8 % / 冗長度: 3.6 % / Biso Wilson estimate: 62.23 Å2 / Rmerge(I) obs: 0.076 / Rsym value: 0.076 / Net I/σ(I): 7.7
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
2.53-2.6
3.6
0.684
1.1
10276
2884
0.684
99.6
2.6-2.67
3.6
0.55
1.4
10106
2824
0.55
99.9
2.67-2.74
3.6
0.46
1.7
9971
2763
0.46
99.7
2.74-2.83
3.6
0.378
2
9552
2653
0.378
99.8
2.83-2.92
3.6
0.285
2
9468
2623
0.285
99.8
2.92-3.02
3.7
0.224
3.2
9210
2522
0.224
99.7
3.02-3.14
3.7
0.17
2.7
8836
2420
0.17
99.9
3.14-3.27
3.7
0.135
5.6
8715
2366
0.135
99.9
3.27-3.41
3.7
0.11
6.7
8366
2264
0.11
100
3.41-3.58
3.7
0.084
8.3
8069
2181
0.084
100
3.58-3.77
3.7
0.068
10.2
7557
2052
0.068
100
3.77-4
3.7
0.058
11.7
7172
1955
0.058
100
4-4.28
3.7
0.047
14.2
6799
1850
0.047
100
4.28-4.62
3.7
0.045
13.8
6300
1724
0.045
100
4.62-5.06
3.6
0.046
13.2
5868
1609
0.046
100
5.06-5.66
3.6
0.045
14.3
5212
1444
0.045
99.9
5.66-6.53
3.6
0.052
12.2
4655
1297
0.052
99.9
6.53-8
3.5
0.054
11.2
3884
1108
0.054
99.9
8-11.31
3.4
0.039
16
2992
878
0.039
99.8
11.31-29.92
3.1
0.036
15.4
1548
500
0.036
94.6
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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
MolProbity
3beta29
モデル構築
SHELX
位相決定
REFMAC
5.2.0019
精密化
SCALA
データスケーリング
PDB_EXTRACT
2
データ抽出
MAR345
CCD
データ収集
MOSFLM
データ削減
CCP4
(SCALA)
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: 2.53→29.921 Å / Cor.coef. Fo:Fc: 0.943 / Cor.coef. Fo:Fc free: 0.927 / SU B: 22.221 / SU ML: 0.235 / TLS residual ADP flag: LIKELY RESIDUAL / 交差検証法: THROUGHOUT / σ(F): 0 / ESU R: 0.648 / ESU R Free: 0.292 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 FOR THE REDUCED SCATTERING POWER DUE TO PARTIAL S-MET INCORPORATION. 4. DENSITY BLOBS OUTSIDE PROTEIN ARE LEFT UNINTEPRETED DUE TO LIMITED RESOLUTION.
Rfactor
反射数
%反射
Selection details
Rfree
0.246
2000
5 %
RANDOM
Rwork
0.214
-
-
-
all
0.215
-
-
-
obs
0.215
39877
99.72 %
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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: BABINET MODEL WITH MASK
原子変位パラメータ
Biso mean: 53.528 Å2
Baniso -1
Baniso -2
Baniso -3
1-
0.31 Å2
0 Å2
0 Å2
2-
-
-0.71 Å2
0 Å2
3-
-
-
0.39 Å2
精密化ステップ
サイクル: LAST / 解像度: 2.53→29.921 Å
タンパク質
核酸
リガンド
溶媒
全体
原子数
7857
0
6
131
7994
拘束条件
Refine-ID
タイプ
Dev ideal
Dev ideal target
数
X-RAY DIFFRACTION
r_bond_refined_d
0.013
0.022
8021
X-RAY DIFFRACTION
r_bond_other_d
0.002
0.02
5260
X-RAY DIFFRACTION
r_angle_refined_deg
1.246
1.961
10844
X-RAY DIFFRACTION
r_angle_other_deg
0.867
3
12864
X-RAY DIFFRACTION
r_dihedral_angle_1_deg
5.042
5
1027
X-RAY DIFFRACTION
r_dihedral_angle_2_deg
32.652
24.399
341
X-RAY DIFFRACTION
r_dihedral_angle_3_deg
16.081
15
1372
X-RAY DIFFRACTION
r_dihedral_angle_4_deg
21.959
15
34
X-RAY DIFFRACTION
r_chiral_restr
0.071
0.2
1243
X-RAY DIFFRACTION
r_gen_planes_refined
0.004
0.02
9014
X-RAY DIFFRACTION
r_gen_planes_other
0.002
0.02
1660
X-RAY DIFFRACTION
r_nbd_refined
0.208
0.2
1535
X-RAY DIFFRACTION
r_nbd_other
0.184
0.2
4938
X-RAY DIFFRACTION
r_nbtor_refined
0.184
0.2
3973
X-RAY DIFFRACTION
r_nbtor_other
0.086
0.2
4429
X-RAY DIFFRACTION
r_xyhbond_nbd_refined
0.145
0.2
196
X-RAY DIFFRACTION
r_xyhbond_nbd_other
0.003
0.2
1
X-RAY DIFFRACTION
r_symmetry_vdw_refined
0.139
0.2
13
X-RAY DIFFRACTION
r_symmetry_vdw_other
0.255
0.2
28
X-RAY DIFFRACTION
r_symmetry_hbond_refined
0.199
0.2
4
X-RAY DIFFRACTION
r_mcbond_it
1.492
3
5458
X-RAY DIFFRACTION
r_mcbond_other
0.45
3
2103
X-RAY DIFFRACTION
r_mcangle_it
2.366
5
8135
X-RAY DIFFRACTION
r_scbond_it
4.023
8
3259
X-RAY DIFFRACTION
r_scangle_it
5.075
11
2705
Refine LS restraints NCS
Dom-ID: 1 / Auth asym-ID: A / Refine-ID: X-RAY DIFFRACTION