- PDB-2mjh: Solution structure of the GLD-1 RNA-binding domain in complex with RNA -
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基本情報
登録情報
データベース: PDB / ID: 2mjh
タイトル
Solution structure of the GLD-1 RNA-binding domain in complex with RNA
要素
5'-CUACUCAUAU-3'
Female germline-specific tumor suppressor gld-1
キーワード
RNA BINDING PROTEIN (RNA結合タンパク質) / GLD-1 / KH-QUA2 domain / STAR protein family (恒星) / RNA regulation / tra-2
機能・相同性
機能・相同性情報
oocyte fate determination / positive regulation of female gonad development / regulation of germ cell proliferation / negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / positive regulation of oocyte development / P granule / regulation of mRNA splicing, via spliceosome / post-transcriptional regulation of gene expression / oogenesis / meiotic cell cycle ...oocyte fate determination / positive regulation of female gonad development / regulation of germ cell proliferation / negative regulation of nuclear-transcribed mRNA catabolic process, nonsense-mediated decay / positive regulation of oocyte development / P granule / regulation of mRNA splicing, via spliceosome / post-transcriptional regulation of gene expression / oogenesis / meiotic cell cycle / mRNA 3'-UTR binding / mRNA 5'-UTR binding / 遺伝子発現の調節 / single-stranded RNA binding / negative regulation of translation / negative regulation of gene expression / mRNA binding / 細胞核 / 細胞質基質 / 細胞質 類似検索 - 分子機能
STAR protein, homodimerisation region / Homodimerisation region of STAR domain protein / KH domain-containing BBP-like / K Homology domain, type 1 / KHドメイン / K Homology domain, type 1 / Ribosomal Protein S8; Chain: A, domain 1 / K Homology domain, type 1 superfamily / KHドメイン / K homology RNA-binding domain ...STAR protein, homodimerisation region / Homodimerisation region of STAR domain protein / KH domain-containing BBP-like / K Homology domain, type 1 / KHドメイン / K Homology domain, type 1 / Ribosomal Protein S8; Chain: A, domain 1 / K Homology domain, type 1 superfamily / KHドメイン / K homology RNA-binding domain / 2-Layer Sandwich / Alpha Beta 類似検索 - ドメイン・相同性
Femalegermline-specifictumorsuppressorgld-1 / Defective in germ line development protein 1
分子量: 16075.546 Da / 分子数: 1 / 断片: KH-QUA2 domain of GLD-1, UNP residues 195-336 / 由来タイプ: 組換発現 由来: (組換発現) Caenorhabditis elegans (センチュウ) 解説: Purification via the IMPACT system (NEB) that results in the protein without any additional residues 遺伝子: gld-1, T23G11.3 / 発現宿主: Escherichia coli (大腸菌) / 株 (発現宿主): BL21(DE3) / 参照: UniProt: Q17339
#2: RNA鎖
5'-CUACUCAUAU-3'
分子量: 3082.868 Da / 分子数: 1 / 由来タイプ: 合成
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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
4
2D 1H-13C HSQC
1
3
2
3D HNCA
1
4
2
3DHN(CO)CA
1
5
2
3D HNCO
1
6
2
3DHN(CA)CO
1
7
2
3DCBCA(CO)NH
1
8
2
3D (H)CCH-TOCSY
1
9
1
3D 1H-15N NOESY
1
10
2
3D 1H-13C NOESY
1
11
3
2D 1H-1H TOCSY
1
12
3
2D 1H-1H NOESY
1
13
3
2D F1f F2f 1H-1H NOESY
1
14
1
2D 1H-13C HSQC
1
15
4
2D F2f 1H-1H NOESY
1
16
4
3D F1f F2e 1H-1H NOESY
1
17
1
Long-range 1H-15N HSQC
1
18
1
3D HNHA
1
19
6
2D 1H-15N IPAPHSQC
1
20
5
2D 1H-13C HSQC
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試料調製
詳細
Solution-ID
内容
溶媒系
1
0.45 mM [U-15N] GLD-1 (aa 195-336), 0.45 mM 5'-CUACUCAUAU-3', 50 mM sodium chloride, 20 mM sodium phosphate, 3 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
2
0.45 mM [U-13C; U-15N] GLD-1 (aa 195-336), 0.45 mM 5'-CUACUCAUAU-3', 50 mM sodium chloride, 20 mM sodium phosphate, 3 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
3
0.45 mM [U-15N] GLD-1 (aa 195-336), 0.45 mM 5'-CUACUCAUAU-3', 50 mM sodium chloride, 20 mM sodium phosphate, 3 mM DTT, 100% D2O
100% D2O
4
0.45 mM [U-13C; U-15N] GLD-1 (aa 195-336), 0.45 mM 5'-CUACUCAUAU-3', 50 mM sodium chloride, 20 mM sodium phosphate, 3 mM DTT, 100% D2O
100% D2O
5
0.45 mM [U-10% 13C; U-100% 15N] GLD-1 (aa 195-336), 0.45 mM 5'-CUACUCAUAU-3', 50 mM sodium chloride, 20 mM sodium phosphate, 3 mM DTT, 90% H2O/10% D2O
90% H2O/10% D2O
6
0.45 mM [U-15N] GLD-1 (aa 195-336), 0.45 mM 5'-CUACUCAUAU-3', 50 mM sodium chloride, 20 mM sodium phosphate, 3 mM DTT, 14 mg/mL Pf1 phage, 90% H2O/10% D2O
90% H2O/10% D2O
試料
濃度 (mg/ml)
構成要素
Isotopic labeling
Solution-ID
0.45mM
GLD-1 (aa 195-336)-1
[U-15N]
1
0.45mM
5'-CUACUCAUAU-3'-2
1
50mM
sodium chloride-3
1
20mM
sodium phosphate-4
1
3mM
DTT-5
1
0.45mM
GLD-1 (aa 195-336)-6
[U-13C; U-15N]
2
0.45mM
5'-CUACUCAUAU-3'-7
2
50mM
sodium chloride-8
2
20mM
sodium phosphate-9
2
3mM
DTT-10
2
0.45mM
GLD-1 (aa 195-336)-11
[U-15N]
3
0.45mM
5'-CUACUCAUAU-3'-12
3
50mM
sodium chloride-13
3
20mM
sodium phosphate-14
3
3mM
DTT-15
3
0.45mM
GLD-1 (aa 195-336)-16
[U-13C; U-15N]
4
0.45mM
5'-CUACUCAUAU-3'-17
4
50mM
sodium chloride-18
4
20mM
sodium phosphate-19
4
3mM
DTT-20
4
0.45mM
GLD-1 (aa 195-336)-21
[U-10% 13C; U-100% 15N]
5
0.45mM
5'-CUACUCAUAU-3'-22
5
50mM
sodium chloride-23
5
20mM
sodium phosphate-24
5
3mM
DTT-25
5
0.45mM
GLD-1 (aa 195-336)-26
[U-15N]
6
0.45mM
5'-CUACUCAUAU-3'-27
6
50mM
sodium chloride-28
6
20mM
sodium phosphate-29
6
3mM
DTT-30
6
14mg/mL
Pf1 phage-31
6
試料状態
イオン強度: 0.17 / pH: 6.5 / 圧: ambient / 温度: 303 K
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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
500
1
Bruker Avance
Bruker
AVANCE
600
2
Bruker Avance
Bruker
AVANCE
700
3
Bruker Avance
Bruker
AVANCE
750
4
Bruker Avance
Bruker
AVANCE
900
5
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解析
NMR software
名称
バージョン
開発者
分類
Amber
9
Case, Darden, Cheatham, III, Simmerling, Wang, Duke, Luo, ... andKollman
NOE constraints total: 2899 / NOE intraresidue total count: 669 / NOE long range total count: 609 / NOE medium range total count: 628 / NOE sequential total count: 841
代表構造
選択基準: closest to the average
NMRアンサンブル
コンフォーマー選択の基準: Lowest energy and NOE violations 計算したコンフォーマーの数: 250 / 登録したコンフォーマーの数: 20