内容: 1 mM [U-100% 13C; U-100% 15N] FOCAL ADHESION KINASE 1, LINKER, 15-MERIC PEPTIDE FROM PAXILLIN-1, 10 mM potassium phosphate-2, 0.1 % sodium azide-3, 5 mM LD2 FROM PAXILLIN-4, 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
1mM
FOCAL ADHESION KINASE 1, LINKER, 15-MERIC PEPTIDE FROM PAXILLIN-1
[U-100% 13C; U-100% 15N]
1
10mM
potassium phosphate-2
1
0.1 %
sodium azide-3
1
5mM
LD2 FROM PAXILLIN-4
1
試料状態
イオン強度: 10 / pH: 6.5 / 圧: AMBIENT Pa / 温度: 310 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
800
1
VARIAN INOVA
Varian
INOVA
600
2
-
解析
NMR software
名称
開発者
分類
CYANA
Guntert, MumenthalerandWuthrich
精密化
CYANA
Guntert, MumenthalerandWuthrich
構造決定
Sparky
Goddard
構造決定
NMRPipe
Delaglio, Grzesiek, Vuister, Zhu, PfeiferandBax
構造決定
MOLMOL
Koradi, BilleterandWuthrich
構造決定
精密化
手法: SIMULATED ANNEALING, TORSION ANGLE DYNAMICS / ソフトェア番号: 1 詳細: THE SAMPLES USED ARE CONSTRUCTS IN WHICH ONE MOTIF IS TETHERED TO THE C-TERMINUS OF THE FAT DOMAIN VIA A GGSX LINKER AND THE OTHER LD MOTIF IS BOUND AS A FREE PEPTIDE. THE GGS LINKERS ARE ...詳細: THE SAMPLES USED ARE CONSTRUCTS IN WHICH ONE MOTIF IS TETHERED TO THE C-TERMINUS OF THE FAT DOMAIN VIA A GGSX LINKER AND THE OTHER LD MOTIF IS BOUND AS A FREE PEPTIDE. THE GGS LINKERS ARE UNSTRUCTURED AND COMPARISON OF SPECTRA LEAD US TO BELIEVE THEY DO NOT IMPOSE ANY CONSTRAINT UPON HOW THE TETHERED LD MOTIFS BIND TO FAT. HOWEVER WHEN ANALYZING THE STRUCTURE IN CYANA, THE PRESENCE OF THE GGS LINKER IN THE SEQUENCE INTERFERS WITH CYANA'S INITIAL ALIGNMENT OF THE 20 LOWEST ENERGY STRUCTURES AND THUS CONFUSES RMSD AND TARGET FUNCTION CALCULATIONS. WE FOUND THAT USING PSEUDO LINKERS IN CYANA RESOLVED THIS PROBLEM WITHOUT CHANGING THE STRUCTURES IN QUESTION AND SO WE CHOSE TO SUBMIT THE STRUCTURES IN THAT WAY. 2RP6 REPRESENTS A COMPOSITE STRUCTURE USING CONSTRAINTS OBTAINED FROM THE 2RP7 AND 2RP9 DATA SETS. IT DOES NOT REPRESENT A NEW CONSTRUCT IN ITSELF, BUT MOST ACCURATELY REPRESENTS THE COMPLETE COMPLEX STRUCTURE BETWEEN PAXILLIN AND FOCAL ADHESION KINASE.
代表構造
選択基準: lowest energy
NMRアンサンブル
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