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基本情報
登録情報 | データベース: PDB / ID: 9b62 | |||||||||
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タイトル | Human RANBP2/RAN(GTP)/RANGAP1-SUMO1/UBC9/CRM1/RAN(GTP) - composite map and model | |||||||||
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![]() | TRANSPORT PROTEIN / karyopherin / SUMO E3 / SUMO E2 / transporter / GTPase / GTPase activating protein / exportin / G-protein | |||||||||
機能・相同性 | ![]() cellular response to triglyceride / cellular response to salt / cellular response to vasopressin / SUMO conjugating enzyme activity / cytoplasmic periphery of the nuclear pore complex / HuR (ELAVL1) binds and stabilizes mRNA / RING-like zinc finger domain binding / SUMO ligase complex / negative regulation of potassium ion transmembrane transporter activity / SUMO ligase activity ...cellular response to triglyceride / cellular response to salt / cellular response to vasopressin / SUMO conjugating enzyme activity / cytoplasmic periphery of the nuclear pore complex / HuR (ELAVL1) binds and stabilizes mRNA / RING-like zinc finger domain binding / SUMO ligase complex / negative regulation of potassium ion transmembrane transporter activity / SUMO ligase activity / protein localization to nuclear pore / : / transferase complex / annulate lamellae / SUMOylation of nuclear envelope proteins / HLH domain binding / SUMO is proteolytically processed / Negative regulation of activity of TFAP2 (AP-2) family transcription factors / SUMO is conjugated to E1 (UBA2:SAE1) / regulation of proteasomal ubiquitin-dependent protein catabolic process / negative regulation of delayed rectifier potassium channel activity / PML body organization / negative regulation of DNA binding / SUMO is transferred from E1 to E2 (UBE2I, UBC9) / negative regulation of action potential / nuclear stress granule / Vitamin D (calciferol) metabolism / pre-miRNA export from nucleus / RNA nuclear export complex / nuclear pore cytoplasmic filaments / mitotic nuclear membrane reassembly / snRNA import into nucleus / synaptonemal complex / regulation of centrosome duplication / nuclear export signal receptor activity / manchette / cellular response to mineralocorticoid stimulus / nuclear inclusion body / small protein activating enzyme binding / Nuclear Pore Complex (NPC) Disassembly / nuclear pore nuclear basket / Transport of Ribonucleoproteins into the Host Nucleus / Regulation of Glucokinase by Glucokinase Regulatory Protein / Defective TPR may confer susceptibility towards thyroid papillary carcinoma (TPC) / Regulation of cholesterol biosynthesis by SREBP (SREBF) / activation of GTPase activity / SUMOylation of DNA methylation proteins / Transport of the SLBP independent Mature mRNA / SUMOylation of immune response proteins / importin-alpha family protein binding / NS1 Mediated Effects on Host Pathways / Transport of the SLBP Dependant Mature mRNA / XY body / NLS-bearing protein import into nucleus / SUMOylation of SUMOylation proteins / regulation of protein export from nucleus / regulation of calcium ion transmembrane transport / Transport of Mature mRNA Derived from an Intronless Transcript / protein localization to nucleolus / negative regulation of protein export from nucleus / Maturation of nucleoprotein / Rev-mediated nuclear export of HIV RNA / nuclear export / Nuclear import of Rev protein / SUMOylation of RNA binding proteins / NEP/NS2 Interacts with the Cellular Export Machinery / kinase activator activity / 転移酵素; アシル基を移すもの; アミノアシル基を移すもの / GTP metabolic process / regulation of cardiac muscle cell contraction / SUMO transferase activity / Transport of Mature mRNA derived from an Intron-Containing Transcript / tRNA processing in the nucleus / Postmitotic nuclear pore complex (NPC) reformation / aggresome / nucleocytoplasmic transport / Maturation of nucleoprotein / centrosome localization / MicroRNA (miRNA) biogenesis / regulation of gluconeogenesis / Viral Messenger RNA Synthesis / DNA metabolic process / negative regulation of protein import into nucleus / SUMOylation of ubiquitinylation proteins / transcription factor binding / ubiquitin-specific protease binding / Vpr-mediated nuclear import of PICs / Maturation of hRSV A proteins / dynein intermediate chain binding / cellular response to cadmium ion / ubiquitin-like protein ligase binding / roof of mouth development / SUMOylation of transcription factors / mitotic sister chromatid segregation / SUMOylation of DNA replication proteins / protein sumoylation / ribosomal large subunit export from nucleus / spermatid development / Estrogen-dependent nuclear events downstream of ESR-membrane signaling / Regulation of HSF1-mediated heat shock response 類似検索 - 分子機能 | |||||||||
生物種 | ![]() | |||||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.9 Å | |||||||||
![]() | Lima, C.D. / DiMattia, M.A. | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structural basis for a nucleoporin exportin complex between RanBP2, SUMO1-RanGAP1, the E2 Ubc9, Crm1 and the Ran GTPase. 著者: Vladimir Baytshtok / Michael A DiMattia / Christopher D Lima / ![]() 要旨: The human nucleoporin RanBP2/Nup358 interacts with SUMO1-modified RanGAP1 and the SUMO E2 Ubc9 at the nuclear pore complex (NPC) to promote export and disassembly of exportin Crm1/Ran(GTP)/cargo ...The human nucleoporin RanBP2/Nup358 interacts with SUMO1-modified RanGAP1 and the SUMO E2 Ubc9 at the nuclear pore complex (NPC) to promote export and disassembly of exportin Crm1/Ran(GTP)/cargo complexes. In mitosis, RanBP2/SUMO1-RanGAP1/Ubc9 remains intact after NPC disassembly and is recruited to kinetochores and mitotic spindles by Crm1 where it contributes to mitotic progression. RanBP2 binds SUMO1-RanGAP1/Ubc9 via motifs that also catalyze SUMO E3 ligase activity. Here, we resolve cryo-EM structures of a RanBP2 C-terminal fragment in complex with Crm1, SUMO1-RanGAP1/Ubc9, and two molecules of Ran(GTP). These structures reveal several interactions with Crm1 including a nuclear export signal (NES) for RanGAP1, the deletion of which mislocalizes RanGAP1 and the Ran GTPase in cells. Our structural and biochemical results support models in which RanBP2 E3 ligase activity is dependent on Crm1, the RanGAP1 NES and Ran GTPase cycling. | |||||||||
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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PDBx/mmCIF形式 | ![]() | 451.2 KB | 表示 | ![]() |
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PDB形式 | ![]() | 349.9 KB | 表示 | ![]() |
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その他 | ![]() |
-検証レポート
文書・要旨 | ![]() | 914.1 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 927.7 KB | 表示 | |
XML形式データ | ![]() | 63.5 KB | 表示 | |
CIF形式データ | ![]() | 97.9 KB | 表示 | |
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-関連構造データ
関連構造データ | ![]() 44235MC C: 同じ文献を引用 ( M: このデータのモデリングに利用したマップデータ |
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類似構造データ | 類似検索 - 機能・相同性 ![]() |
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リンク
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集合体
登録構造単位 | ![]()
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要素
-タンパク質 , 6種, 7分子 ABFCDEG
#1: タンパク質 | 分子量: 123874.781 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: SHM N-terminal amino acids remnants of expression tag 由来: (組換発現) ![]() ![]() ![]() | ||||||||
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#2: タンパク質 | 分子量: 25012.764 Da / 分子数: 2 / Mutation: Q69L / 由来タイプ: 組換発現 詳細: GSHMAS N-terminal amino acids residual after removal of purification epitope tag 由来: (組換発現) ![]() ![]() ![]() 参照: UniProt: P62826, 加水分解酵素; 酸無水物に作用; GTPに作用・細胞または細胞小器官の運動に関与 #3: タンパク質 | | 分子量: 18313.092 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: GSH N-terminal amino acids left over after cleavage of affinity tag 由来: (組換発現) ![]() ![]() ![]() 参照: UniProt: P63279, 転移酵素; アシル基を移すもの; アミノアシル基を移すもの #4: タンパク質 | | 分子量: 11532.003 Da / 分子数: 1 / Mutation: Q94P / 由来タイプ: 組換発現 詳細: GSHM N-terminal amino acids left over after cleavage of affinity tag 由来: (組換発現) ![]() ![]() ![]() #5: タンパク質 | | 分子量: 68399.422 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: GSH N-terminal amino acids left after cleavage of affinity tag The residues that are misaligned in your alignment should be aligned to residue 66 with a gap after. please fix 由来: (組換発現) ![]() ![]() ![]() 参照: UniProt: P49792, 転移酵素; アシル基を移すもの; アミノアシル基を移すもの #6: タンパク質 | | 分子量: 64022.172 Da / 分子数: 1 / 由来タイプ: 組換発現 詳細: GSHM N-terminal amino acids left after cleavage of affinity tag 由来: (組換発現) ![]() ![]() ![]() |
-非ポリマー , 2種, 4分子 


#7: 化合物 | #8: 化合物 | |
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-詳細
研究の焦点であるリガンドがあるか | N |
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Has protein modification | N |
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
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試料調製
構成要素 | 名称: Human RANBP2/RAN(GTP)/RANGAP1-SUMO1/UBC9/CRM1/RAN(GTP) complex タイプ: COMPLEX 詳細: SUMO1 Gly97 is covalently linked to RANGAP1 Lys524 through a isopeptide bond Entity ID: #1-#6 / 由来: RECOMBINANT |
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分子量 | 実験値: NO |
由来(天然) | 生物種: ![]() |
由来(組換発現) | 生物種: ![]() ![]() |
緩衝液 | pH: 8 詳細: 20 mM Tris-Cl pH 8.0, 50 mM NaCl, 0.1 mM TCEP supplemented with 0.02% (v/v) IGEPAL CA-630 |
試料 | 濃度: 8 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: Gel filtered - sample was monodisperse |
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 295 K / 詳細: 30 s wait time, blot for 2.5 s before plunging |
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電子顕微鏡撮影
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: FEI TITAN KRIOS |
電子銃 | 電子線源: ![]() |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 3000 nm / 最小 デフォーカス(公称値): 1000 nm / Cs: 2.7 mm |
試料ホルダ | 凍結剤: NITROGEN 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER |
撮影 | 平均露光時間: 10 sec. / 電子線照射量: 85.2 e/Å2 / 検出モード: SUPER-RESOLUTION フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) |
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解析
EMソフトウェア |
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CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | |||||||||||||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 895509 詳細: This is a composite map so numbers of particles per map component differs - after selection of particles containing Crm1 there were 704835 unique particles - after selection of particles ...詳細: This is a composite map so numbers of particles per map component differs - after selection of particles containing Crm1 there were 704835 unique particles - after selection of particles containing Crm1-Ran(GTP) there were 534708 unique particles | |||||||||||||||||||||||||||||||||||
3次元再構成 | 解像度: 2.9 Å / 解像度の算出法: OTHER / 粒子像の数: 534708 詳細: Composite map - estimated best average resolution of FR maps 2.9 Related particles and maps used to generate the composite listed in Related Entries 対称性のタイプ: POINT | |||||||||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: OTHER / 空間: REAL 詳細: Atomic model was manually fit and refined using composite map, and manually inspected using Coot | |||||||||||||||||||||||||||||||||||
原子モデル構築 |
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拘束条件 |
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