+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 6r6b | ||||||||||||||||||
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タイトル | Structure of the core Shigella flexneri type III secretion system export gate complex SctRST (Spa24/Spa9/Spa29). | ||||||||||||||||||
要素 |
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キーワード | PROTEIN TRANSPORT / Type III secretion / T3SS / Flagella (鞭毛) / Cryo-EM (低温電子顕微鏡法) / Membrane protein (膜タンパク質) | ||||||||||||||||||
機能・相同性 | 機能・相同性情報 | ||||||||||||||||||
生物種 | Shigella flexneri (フレクスナー赤痢菌) | ||||||||||||||||||
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: 3.5 Å | ||||||||||||||||||
データ登録者 | Johnson, S. / Kuhlen, L. / Deme, J.C. / Abrusci, P. / Lea, S.M. | ||||||||||||||||||
資金援助 | 英国, 5件
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引用 | ジャーナル: mBio / 年: 2019 タイトル: The Structure of an Injectisome Export Gate Demonstrates Conservation of Architecture in the Core Export Gate between Flagellar and Virulence Type III Secretion Systems. 著者: Steven Johnson / Lucas Kuhlen / Justin C Deme / Patrizia Abrusci / Susan M Lea / 要旨: Export of proteins through type III secretion systems (T3SS) is critical for motility and virulence of many major bacterial pathogens. Proteins are exported through a genetically defined export gate ...Export of proteins through type III secretion systems (T3SS) is critical for motility and virulence of many major bacterial pathogens. Proteins are exported through a genetically defined export gate complex consisting of three proteins. We have recently shown at 4.2 Å that the flagellar complex of these three putative membrane proteins (FliPQR in flagellar systems, SctRST in virulence systems) assembles into an extramembrane helical assembly that likely seeds correct assembly of the rod. Here we present the structure of an equivalent complex from the virulence system at 3.5 Å by cryo-electron microscopy. This higher-resolution structure yields a more precise description of the structure and confirms the prediction of structural conservation in this core complex. Analysis of particle heterogeneity also suggests how the SctS/FliQ subunits sequentially assemble in the complex. Although predicted on the basis of sequence conservation, the work presented here formally demonstrates that all classes of type III secretion systems, flagellar or virulence, share the same architecture at the level of the core structures. This absolute conservation of the unusual extramembrane structure of the core export gate complex now allows work to move to focusing on both mechanistic studies of type III but also on fundamental studies of how such a complex is assembled. #1: ジャーナル: Nat Struct Mol Biol / 年: 2018 タイトル: Structure of the core of the type III secretion system export apparatus. 著者: Lucas Kuhlen / Patrizia Abrusci / Steven Johnson / Joseph Gault / Justin Deme / Joseph Caesar / Tobias Dietsche / Mehari Tesfazgi Mebrhatu / Tariq Ganief / Boris Macek / Samuel Wagner / Carol ...著者: Lucas Kuhlen / Patrizia Abrusci / Steven Johnson / Joseph Gault / Justin Deme / Joseph Caesar / Tobias Dietsche / Mehari Tesfazgi Mebrhatu / Tariq Ganief / Boris Macek / Samuel Wagner / Carol V Robinson / Susan M Lea / 要旨: Export of proteins through type III secretion systems is critical for motility and virulence of many major bacterial pathogens. Three putative integral membrane proteins (FliP, FliQ, FliR) are ...Export of proteins through type III secretion systems is critical for motility and virulence of many major bacterial pathogens. Three putative integral membrane proteins (FliP, FliQ, FliR) are suggested to form the core of an export gate in the inner membrane, but their structure, assembly and location within the final nanomachine remain unclear. Here, we present the cryoelectron microscopy structure of the Salmonella Typhimurium FliP-FliQ-FliR complex at 4.2 Å. None of the subunits adopt canonical integral membrane protein topologies, and common helix-turn-helix structural elements allow them to form a helical assembly with 5:4:1 stoichiometry. Fitting of the structure into reconstructions of intact secretion systems, combined with cross-linking, localize the export gate as a core component of the periplasmic portion of the machinery. This study thereby identifies the export gate as a key element of the secretion channel and implies that it primes the helical architecture of the components assembling downstream. | ||||||||||||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | 分子: MolmilJmol/JSmol |
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 6r6b.cif.gz | 255.3 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb6r6b.ent.gz | 210.7 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 6r6b.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/r6/6r6b ftp://data.pdbj.org/pub/pdb/validation_reports/r6/6r6b | HTTPS FTP |
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-関連構造データ
-リンク
-集合体
登録構造単位 |
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-要素
#1: タンパク質 | 分子量: 24215.562 Da / 分子数: 5 / 由来タイプ: 組換発現 由来: (組換発現) Shigella flexneri (フレクスナー赤痢菌) 遺伝子: spaP, spa24, CP0153 / 発現宿主: Escherichia coli (大腸菌) / Variant (発現宿主): MT56 / 参照: UniProt: P0A1L3 #2: タンパク質 | | 分子量: 32644.086 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) Shigella flexneri (フレクスナー赤痢菌) 遺伝子: spaR, spa29, CP0155 / 発現宿主: Escherichia coli (大腸菌) / Variant (発現宿主): MT56 / 参照: UniProt: P0A1M6 #3: タンパク質 | 分子量: 9433.338 Da / 分子数: 4 / 由来タイプ: 組換発現 由来: (組換発現) Shigella flexneri (フレクスナー赤痢菌) 遺伝子: spaQ, spa9, CP0154 / 発現宿主: Escherichia coli (大腸菌) / Variant (発現宿主): MT56 / 参照: UniProt: P0A1M4 |
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-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 |
-試料調製
構成要素 | 名称: Shigella flexneri type III secretion export gate (Spa24Spa9Spa29)Type three secretion system タイプ: COMPLEX / Entity ID: all / 由来: RECOMBINANT | ||||||||||||||||
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分子量 | 値: 0.191 MDa / 実験値: YES | ||||||||||||||||
由来(天然) | 生物種: Shigella flexneri (フレクスナー赤痢菌) | ||||||||||||||||
由来(組換発現) | 生物種: Escherichia coli (大腸菌) / 株: MT56 | ||||||||||||||||
緩衝液 | pH: 8 | ||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 8.4 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES | ||||||||||||||||
試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: Quantifoil, UltrAuFoil, R1.2/1.3 | ||||||||||||||||
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 |
-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: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: SPOT SCAN |
電子レンズ | モード: BRIGHT FIELDBright-field microscopy |
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: 8 sec. / 電子線照射量: 48 e/Å2 / 検出モード: COUNTING フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 実像数: 3741 |
画像スキャン | 動画フレーム数/画像: 20 |
-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: 775073 | ||||||||||||||||||||||||||||
対称性 | 点対称性: C1 (非対称) | ||||||||||||||||||||||||||||
3次元再構成 | 解像度: 3.5 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 212561 / 対称性のタイプ: POINT | ||||||||||||||||||||||||||||
原子モデル構築 | プロトコル: AB INITIO MODEL / 空間: REAL |