ジャーナル: Febs J. / 年: 2016 タイトル: Structural studies reveal an important role for the pleiotrophin C-terminus in mediating interactions with chondroitin sulfate. 著者: Ryan, E. / Shen, D. / Wang, X.
分子量: 15346.931 Da / 分子数: 1 / 断片: UNP residues 33-168 / 由来タイプ: 組換発現 / 詳細: Two domains connected by a flexible linker / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: HBNF1, NEGF1, PTN / 発現宿主: Escherichia coli (大腸菌) / 株 (発現宿主): Origami BL21(DE3) / 参照: UniProt: P21246
Has protein modification
Y
-
実験情報
-
実験
実験
手法: 溶液NMR / 詳細: Solution Structure of Pleiotrophin
NMR実験
Conditions-ID
Experiment-ID
Solution-ID
タイプ
1
1
1
2D 1H-15N HSQC
1
2
1
3D HN(CA)CB
1
3
1
3DCBCA(CO)NH
1
4
1
3DH(CCO)NH
1
5
1
3D (H)CCH-TOCSY
1
6
1
3D 1H-15N NOESY
1
7
1
3D 1H-13C NOESY
1
8
1
3D 1H-13C NOESY
-
試料調製
詳細
内容: 0.6 mM [U-100% 13C; U-100% 15N] PTN, 10 % v/v D2O, 10 mM MES, 150 mM sodium chloride, 90% H2O/10% D2O 溶媒系: 90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.6mM
PTN-1
[U-100% 13C; U-100% 15N]
1
10v/v
D2O-2
1
10mM
MES-3
1
150mM
sodium chloride-4
1
試料状態
イオン強度: 0.15 / pH: 6 / 圧: ambient / 温度: 298 K
-
NMR測定
NMRスペクトロメーター
タイプ
製造業者
モデル
磁場強度 (MHz)
Spectrometer-ID
Bruker Avance
Bruker
AVANCE
600
1
Varian INOVA
Varian
INOVA
800
2
-
解析
NMR software
名称
開発者
分類
NMRPipe
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
解析
NMRView
Johnson, OneMoonScientific
データ解析
CYANA
Guntert, MumenthalerandWuthrich
構造決定
X-PLOR NIH
Schwieters, Kuszewski, TjandraandClore
精密化
精密化
手法: simulated annealing / ソフトェア番号: 1 / 詳細: water refinement with the default protocol
NMR constraints
NOE constraints total: 749 / NOE intraresidue total count: 263 / NOE long range total count: 147 / NOE medium range total count: 44 / NOE sequential total count: 295 / Protein chi angle constraints total count: 0 / Protein other angle constraints total count: 0 / Protein phi angle constraints total count: 63 / Protein psi angle constraints total count: 62
代表構造
選択基準: fewest violations
NMRアンサンブル
Average torsion angle constraint violation: 0.9 ° コンフォーマー選択の基準: structures with the least restraint violations 計算したコンフォーマーの数: 100 / 登録したコンフォーマーの数: 10 / Maximum lower distance constraint violation: 0 Å / Maximum torsion angle constraint violation: 9 ° / Maximum upper distance constraint violation: 0.73 Å