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基本情報
登録情報 | データベース: PDB / ID: 6mrt | ||||||
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タイトル | 12-meric ClyA pore complex | ||||||
![]() | Hemolysin E, chromosomal | ||||||
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機能・相同性 | ![]() modulation of apoptotic process in another organism / ![]() ![]() ![]() ![]() 類似検索 - 分子機能 | ||||||
生物種 | ![]() ![]() ![]() | ||||||
手法 | ![]() ![]() ![]() | ||||||
![]() | Peng, W. / de Souza Santos, M. / Li, Y. / Tomchick, D.R. / Orth, K. | ||||||
![]() | ![]() タイトル: High-resolution cryo-EM structures of the E. coli hemolysin ClyA oligomers. 著者: Wei Peng / Marcela de Souza Santos / Yang Li / Diana R Tomchick / Kim Orth / ![]() 要旨: Pore-forming proteins (PFPs) represent a functionally important protein family, that are found in organisms from viruses to humans. As a major branch of PFPs, bacteria pore-forming toxins (PFTs) ...Pore-forming proteins (PFPs) represent a functionally important protein family, that are found in organisms from viruses to humans. As a major branch of PFPs, bacteria pore-forming toxins (PFTs) permeabilize membranes and usually cause the death of target cells. E. coli hemolysin ClyA is the first member with the pore complex structure solved among α-PFTs, employing α-helices as transmembrane elements. ClyA is proposed to form pores composed of various numbers of protomers. With high-resolution cryo-EM structures, we observe that ClyA pore complexes can exist as newly confirmed oligomers of a tridecamer and a tetradecamer, at estimated resolutions of 3.2 Å and 4.3 Å, respectively. The 2.8 Å cryo-EM structure of a dodecamer dramatically improves the existing structural model. Structural analysis indicates that protomers from distinct oligomers resemble each other and neighboring protomers adopt a conserved interaction mode. We also show a stabilized intermediate state of ClyA during the transition process from soluble monomers to pore complexes. Unexpectedly, even without the formation of mature pore complexes, ClyA can permeabilize membranes and allow leakage of particles less than ~400 Daltons. In addition, we are the first to show that ClyA forms pore complexes in the presence of cholesterol within artificial liposomes. These findings provide new mechanistic insights into the dynamic process of pore assembly for the prototypical α-PFT ClyA. | ||||||
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構造の表示
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構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
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PDBx/mmCIF形式 | ![]() | 783.1 KB | 表示 | ![]() |
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PDB形式 | ![]() | 622.8 KB | 表示 | ![]() |
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-関連構造データ
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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
#1: タンパク質 | 分子量: 36131.816 Da / 分子数: 12 / 由来タイプ: 組換発現 由来: (組換発現) ![]() ![]() ![]() 株: K12 / 遺伝子: hlyE, clyA, hpr, sheA, ycgD, b1182, JW5181 / 発現宿主: ![]() ![]() ![]() |
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-実験情報
-実験
実験 | 手法: ![]() |
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EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: ![]() |
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試料調製
構成要素 | 名称: 12-meric ClyA pore complex / タイプ: COMPLEX / Entity ID: all / 由来: RECOMBINANT |
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分子量 | 値: 0.41 MDa / 実験値: YES |
由来(天然) | 生物種: ![]() ![]() ![]() |
由来(組換発現) | 生物種: ![]() ![]() ![]() |
緩衝液 | pH: 8 |
試料 | 包埋: NO / シャドウイング: NO / 染色![]() ![]() |
試料支持 | 詳細: unspecified |
急速凍結![]() | 凍結剤: ETHANE |
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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![]() |
撮影 | 電子線照射量: 50 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
CTF補正![]() | タイプ: NONE |
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3次元再構成![]() | 解像度: 2.8 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 482946 / 対称性のタイプ: POINT |