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基本情報
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タイトル | Structure of the rabbit 80S ribosome stalled on a 4-TMD Rhodopsin intermediate in complex with the multipass translocon | ||||||||||||
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![]() | Ribosome / Membrane protein / translocon | ||||||||||||
機能・相同性 | ![]() multi-pass transmembrane protein insertion into ER membrane / multi-pass translocon complex / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / endoplasmic reticulum Sec complex / pronephric nephron development / protein localization to nuclear inner membrane / rough endoplasmic reticulum membrane / cotranslational protein targeting to membrane / Sec61 translocon complex / protein targeting to ER ...multi-pass transmembrane protein insertion into ER membrane / multi-pass translocon complex / Insertion of tail-anchored proteins into the endoplasmic reticulum membrane / endoplasmic reticulum Sec complex / pronephric nephron development / protein localization to nuclear inner membrane / rough endoplasmic reticulum membrane / cotranslational protein targeting to membrane / Sec61 translocon complex / protein targeting to ER / protein insertion into ER membrane / protein-transporting ATPase activity / SRP-dependent cotranslational protein targeting to membrane, translocation / signal sequence binding / post-translational protein targeting to membrane, translocation / endoplasmic reticulum calcium ion homeostasis / ribosomal subunit / protein folding chaperone complex / ER overload response / ubiquitin ligase inhibitor activity / protein transmembrane transporter activity / positive regulation of signal transduction by p53 class mediator / regulation of signal transduction / protein-RNA complex assembly / rough endoplasmic reticulum / ERAD pathway / MDM2/MDM4 family protein binding / protein folding chaperone / cytosolic ribosome / guanyl-nucleotide exchange factor activity / maturation of LSU-rRNA from tricistronic rRNA transcript (SSU-rRNA, 5.8S rRNA, LSU-rRNA) / ribosomal large subunit biogenesis / post-embryonic development / antimicrobial humoral immune response mediated by antimicrobial peptide / large ribosomal subunit / heparin binding / regulation of translation / ribosome binding / 5S rRNA binding / ribosomal large subunit assembly / large ribosomal subunit rRNA binding / nuclear membrane / defense response to Gram-negative bacterium / cytosolic large ribosomal subunit / killing of cells of another organism / cytoplasmic translation / tRNA binding / postsynaptic density / rRNA binding / structural constituent of ribosome / ribosome / translation / ribonucleoprotein complex / mRNA binding / synapse / calcium ion binding / endoplasmic reticulum membrane / nucleolus / endoplasmic reticulum / RNA binding / zinc ion binding / nucleus / membrane / plasma membrane / cytosol / cytoplasm 類似検索 - 分子機能 | ||||||||||||
生物種 | ![]() ![]() ![]() ![]() | ||||||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.88 Å | ||||||||||||
![]() | Kim MK / Lewis AJO / Keenan RJ / Hegde RS | ||||||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Mechanism of an intramembrane chaperone for multipass membrane proteins. 著者: Luka Smalinskaitė / Min Kyung Kim / Aaron J O Lewis / Robert J Keenan / Ramanujan S Hegde / ![]() ![]() 要旨: Multipass membrane proteins play numerous roles in biology and include receptors, transporters, ion channels and enzymes. How multipass proteins are co-translationally inserted and folded at the ...Multipass membrane proteins play numerous roles in biology and include receptors, transporters, ion channels and enzymes. How multipass proteins are co-translationally inserted and folded at the endoplasmic reticulum is not well understood. The prevailing model posits that each transmembrane domain (TMD) of a multipass protein successively passes into the lipid bilayer through a front-side lateral gate of the Sec61 protein translocation channel. The PAT complex, an intramembrane chaperone comprising Asterix and CCDC47, engages early TMDs of multipass proteins to promote their biogenesis by an unknown mechanism. Here, biochemical and structural analysis of intermediates during multipass protein biogenesis showed that the nascent chain is not engaged with Sec61, which is occluded and latched closed by CCDC47. Instead, Asterix binds to and redirects the substrate to a location behind Sec61, where the PAT complex contributes to a multipass translocon surrounding a semi-enclosed, lipid-filled cavity. Detection of multiple TMDs in this cavity after their emergence from the ribosome suggests that multipass proteins insert and fold behind Sec61. Accordingly, biogenesis of several multipass proteins was unimpeded by inhibitors of the Sec61 lateral gate. These findings elucidate the mechanism of an intramembrane chaperone and suggest a new framework for multipass membrane protein biogenesis at the endoplasmic reticulum. | ||||||||||||
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構造の表示
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-EMDBアーカイブ
マップデータ | ![]() | 218.7 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 69.2 KB 69.2 KB | 表示 表示 | ![]() |
画像 | ![]() | 87.4 KB | ||
Filedesc metadata | ![]() | 15.9 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 554.5 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 554.1 KB | 表示 | |
XML形式データ | ![]() | 7.5 KB | 表示 | |
CIF形式データ | ![]() | 8.6 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 | ![]() 7tutMC ![]() 7tm3C C: 同じ文献を引用 ( M: このマップから作成された原子モデル |
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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
EMDBのページ | ![]() ![]() |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||
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注釈 | Reference map | ||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.34 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
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試料の構成要素
+全体 : 80S ribosome translating a stalled, four-TMD nascent chain (deriv...
+超分子 #1: 80S ribosome translating a stalled, four-TMD nascent chain (deriv...
+分子 #1: uL2
+分子 #2: uL4
+分子 #3: uL18
+分子 #4: eL6
+分子 #5: uL30
+分子 #6: eL8
+分子 #7: uL6
+分子 #8: uL16
+分子 #9: uL5
+分子 #10: eL13
+分子 #11: eL14
+分子 #12: eL15
+分子 #13: uL13
+分子 #14: uL22
+分子 #15: eL18
+分子 #16: eL19
+分子 #17: eL20
+分子 #18: eL21
+分子 #19: eL22
+分子 #20: uL14
+分子 #21: eL24
+分子 #22: eL23
+分子 #23: uL24
+分子 #24: eL27
+分子 #25: uL15
+分子 #26: eL29
+分子 #27: eL30
+分子 #28: eL31
+分子 #29: eL32
+分子 #30: eL33
+分子 #31: eL34
+分子 #32: eL35
+分子 #33: eL36
+分子 #34: eL37
+分子 #35: eL38
+分子 #36: eL39
+分子 #37: eL40
+分子 #38: eL41
+分子 #39: eL42
+分子 #40: eL43
+分子 #42: eL28
+分子 #45: uL3
+分子 #46: Nascent chain
+分子 #47: Protein transport protein Sec61 subunit alpha isoform 1
+分子 #48: Protein transport protein Sec61 subunit beta
+分子 #49: Protein transport protein Sec61 gamma
+分子 #50: Coiled-coil domain containing 47
+分子 #51: PAT complex subunit Asterix
+分子 #52: Nicalin
+分子 #53: Transmembrane protein 147
+分子 #54: Calcium load-activated calcium channel
+分子 #56: Obligate partner of TMCO1 insertase
+分子 #41: P-site tRNA
+分子 #43: 5S ribosomal RNA
+分子 #44: 5.8S ribosomal RNA
+分子 #55: 28S ribosomal RNA
+分子 #57: MAGNESIUM ION
+分子 #58: ZINC ION
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
緩衝液 | pH: 7.5 詳細: 50 mM HEPES-KOH, pH 7.5, 200 mM potassium acetate, 5 mM magnesium acetate, 0.25% digitonin, 50 mM biotin |
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凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: FEI VITROBOT MARK IV 詳細: Blot for 4 seconds with Whatman filter papers at a blot force of -15 before plunging.. |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 撮影したグリッド数: 1 / 実像数: 13755 / 平均電子線量: 54.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2.7 µm 最小 デフォーカス(公称値): 1.9000000000000001 µm |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |
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画像解析
初期モデル | モデルのタイプ: OTHER |
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最終 再構成 | 解像度のタイプ: BY AUTHOR / 解像度: 3.88 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 使用した粒子像数: 136812 |
初期 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
最終 角度割当 | タイプ: MAXIMUM LIKELIHOOD |
-原子モデル構築 1
精密化 | 空間: REAL / プロトコル: FLEXIBLE FIT |
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得られたモデル | ![]() PDB-7tut: |