AUTHORS STATE THAT THE ISOTOPICALLY LABELLED BIOSYNTHETIC PEPTIDES HAVE GS CLONING ARTIFACT AT THE ...AUTHORS STATE THAT THE ISOTOPICALLY LABELLED BIOSYNTHETIC PEPTIDES HAVE GS CLONING ARTIFACT AT THE N-TERMINI, WHICH WERE USED PRIMARILY TO DERIVE THE DATA THAT LEAD TO THE RELEVANT PARTS OF THE STRUCTURE. HOWEVER, THE NATURAL ISOTOPIC ABUNDANCE SYNTHETIC PEPTIDES WERE ALSO USED FOR SOME NMR EXPERIMENTS. THEY DO NOT HAVE GS CLONING ARTIFACT. INSTEAD, THEY HAVE ACETYL GROUP AT THE N-TERMINUS AND AMIDE AT THE C-TERMINUS.
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実験情報
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実験
実験
手法: 溶液NMR
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
2D 1H-15N HSQC
1
2
1
2D 1H-15N HSQC
1
3
1
2D 1H-15N HSQC
1
4
1
2D 1H-15N HSQC
1
5
2
2D 1H-15N HSQC
1
6
2
2D 1H-15N HSQC
1
7
2
2D 1H-15N HSQC
1
8
2
2D 1H-15N HSQC
1
9
3
2D 1H-15N HSQC
1
10
3
2D 1H-15N HSQC
1
11
3
2D 1H-15N HSQC
1
12
3
2D 1H-15N HSQC
1
13
1
3D HNCO
1
14
1
3D HNCA
1
15
2
3D HNCA
1
16
3
3D HNCA
1
17
4
3D HN(CA)CB
1
18
1
3DHN(CO)CA
1
19
2
3DHN(CO)CA
1
20
3
3DHN(CO)CA
1
21
1
3D (H)CCH-TOCSY
1
22
2
3D (H)CCH-TOCSY
1
23
3
3D (H)CCH-TOCSY
1
24
1
3D 1H-15N NOESY
1
25
2
3D 1H-15N NOESY
1
26
3
3D 1H-15N NOESY
1
27
5
3D 1H-15N NOESY
1
28
1
3D 1H-13C NOESY aliphatic
1
29
2
3D 1H-13C NOESY aliphatic
1
30
3
3D 1H-13C NOESY aliphatic
1
31
5
3D 1H-13C NOESY aliphatic
1
32
1
3D 1H-13C NOESY aromatic
1
33
2
3D 1H-13C NOESY aromatic
1
34
3
3D 1H-13C NOESY aromatic
1
35
5
3D (H)CCH-TOCSY
1
36
1
3D 1H-13C NOESY filtered/edited
1
37
2
3D 1H-13C NOESY filtered/edited
1
38
1
(HB)CB(CGCD)HD
1
39
1
3D CCH-TOCSY
1
40
3
3DCBCA(CO)NH
1
41
3
3D 1H-15N NOESY ARG-centered
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試料調製
詳細
Solution-ID
内容
溶媒系
1
25 mM sodium phosphate, 250 mM NaCl, 1 mM sodium azide, 0.9-1.3 mM [U-13C; U-15N] Snu17p, 1.1-1.5 mM Bud13p, 1.4-1.9 mM Pml1p, 90% H2O/10% D2O
90% H2O/10% D2O
2
25 mM sodium phosphate, 250 mM NaCl, 1 mM sodium azide, 1.0-1.3 mM Snu17p, 1.2-1.5 mM Bud13p, 0.8-1.0 mM [U-13C; U-15N] Pml1p, 90% H2O/10% D2O
90% H2O/10% D2O
3
25 mM sodium phosphate, 250 mM NaCl, 1 mM sodium azide, 1.0-1.3 mM Snu17p, 0.8-1.0 mM [U-10% 13C; U-99% 15N] Bud13p, 1.5-1.9 mM Pml1p, 90% H2O/10% D2O
90% H2O/10% D2O
4
25 mM sodium phosphate, 250 mM NaCl, 1 mM sodium azide, 0.9-1.3 mM [U-13C; U-15N; U-2H] Snu17p, 1.1-1.5 mM Bud13p, 1.4-1.9 mM Pml1p, 90% H2O/10% D2O
90% H2O/10% D2O
5
25 mM sodium phosphate, 250 mM NaCl, 1 mM sodium azide, 1.0-1.3 mM [U-13C; U-15N; U-2H] Snu17p, 0.8-1.0 mM [U-10% 13C; U-99% 15N] Bud13p, 1.5-1.9 mM Pml1p, 90% H2O/10% D2O
NOE constraints total: 4105 / NOE intraresidue total count: 769 / NOE long range total count: 1597 / NOE medium range total count: 775 / NOE sequential total count: 964 / Hydrogen bond constraints total count: 50 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 0 / Protein phi angle constraints total count: 115 / Protein psi angle constraints total count: 115
代表構造
選択基準: closest to the average
NMRアンサンブル
Average torsion angle constraint violation: 0.62 ° / コンフォーマー選択の基準: target function / 計算したコンフォーマーの数: 500 / 登録したコンフォーマーの数: 20 / Maximum lower distance constraint violation: 0 Å / Maximum torsion angle constraint violation: 3.2 ° / Maximum upper distance constraint violation: 0.307 Å / Torsion angle constraint violation method: CNS
NMR ensemble rms
Distance rms dev: 0.0101 Å / Distance rms dev error: 0.0008 Å