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基本情報
登録情報 | データベース: PDB / ID: 8hfs | ||||||
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タイトル | The structure of LcnA, LciA, and the man-PTS of Lactococcus lactis | ||||||
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![]() | MEMBRANE PROTEIN / antibiotic resistance / antimicrobial peptides / mannose phosphotransferase system / man-PTS / bacteriocins / non-pediocin-like/class IId bacteriocins / lactococcin A / LcnA / immunity / LciA / self-protection | ||||||
機能・相同性 | ![]() protein-Npi-phosphohistidine-sugar phosphotransferase / protein-N(pi)-phosphohistidine--N-acetyl-D-glucosamine phosphotransferase activity / bacteriocin immunity / cytolysis / phosphoenolpyruvate-dependent sugar phosphotransferase system / defense response to bacterium / host cell plasma membrane / extracellular region / membrane / plasma membrane 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: 2.98 Å | ||||||
![]() | Wang, J.W. | ||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Structural Basis of the Mechanisms of Action and Immunity of Lactococcin A, a Class IId Bacteriocin. 著者: Ruilian Li / Jinsong Duan / Yicheng Zhou / Jiawei Wang / ![]() ![]() 要旨: Lactococcin A (LcnA), a class IId bacteriocin, induces membrane leakage and cell death by specifically binding to the membrane receptor-mannose phosphotransferase system (man-PTS), as is the case for ...Lactococcin A (LcnA), a class IId bacteriocin, induces membrane leakage and cell death by specifically binding to the membrane receptor-mannose phosphotransferase system (man-PTS), as is the case for pediocin-like (class IIa) bacteriocins. The cognate immunity protein of bacteriocins, which protects the producer cell from its own bacteriocin, recognizes and binds to the bacteriocin-man-PTS complex, consequently blocking membrane leakage. We previously deciphered the mode of action and immunity of class IIa bacteriocins. Here, we determined the structure of the ternary complex of LcnA, LciA (, the immunity protein), and its receptor, , the man-PTS of Lactococcus lactis (ll-man-PTS). An external loop on the membrane-located component IIC of ll-man-PTS was found to prevent specific binding of the N-terminal region of LcnA to the site recognized by pediocin-like bacteriocins. Thus, the N-terminal β-sheet region of LcnA recognized an adjacent site on the extracellular side of ll-man-PTS, with the LcnA C-terminal hydrophobic helix penetrating into the membrane. The cytoplasmic cleft formed within the man-PTS Core and Vmotif domains induced by embedded LcnA from the periplasmic side is adopted by the appropriate angle between helices H3 and H4 of the N terminus of LciA. The flexible C terminus of LciA then blocks membrane leakage. To summarize, our findings reveal the molecular mechanisms of action and immunity of LcnA and LciA, laying a foundation for further design of class IId bacteriocins. Class IId (lactococcin-like) bacteriocins and class IIa (pediocin-like) bacteriocins share a few similarities: (i) both induce membrane leakage and cell death by specifically binding the mannose phosphotransferase system (man-PTS) on their target cells, and (ii) cognate immunity proteins recognize and bind to the bacteriocin-man-PTS complex to block membrane leakage. However, class IId bacteriocins lack the "pediocin box" motif, which is typical of class IIa bacteriocins, and basically target only lactococcal cells; in contrast, class IIa bacteriocins target diverse bacterial cells, but not lactococcal cells. We previously solved the structure of class IIa bacteriocin-receptor-immunity ternary complex from Lactobacillus sakei. Here, we determined the structure of the ternary complex of class IId bacteriocin LcnA, its cognate immunity protein LciA, and its receptor, the man-PTS of Lactococcus lactis. By comparing the interactions between man-PTS and class IIa and class IId bacteriocins, this study affords some clues to better understand the specificity of bacteriocins targeting the mannose phosphotransferase system. | ||||||
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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-検証レポート
文書・要旨 | ![]() | 1.1 MB | 表示 | ![]() |
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文書・詳細版 | ![]() | 1.1 MB | 表示 | |
XML形式データ | ![]() | 62.4 KB | 表示 | |
CIF形式データ | ![]() | 98.3 KB | 表示 | |
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-関連構造データ
関連構造データ | ![]() 34726MC M: このデータのモデリングに利用したマップデータ C: 同じ文献を引用 ( |
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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
#1: タンパク質 | 分子量: 27572.371 Da / 分子数: 3 / 由来タイプ: 組換発現 由来: (組換発現) ![]() 遺伝子: ptnC, LLKF_1869 / 発現宿主: ![]() ![]() 参照: UniProt: D2BKY8, protein-Npi-phosphohistidine-sugar phosphotransferase #2: タンパク質 | 分子量: 33630.473 Da / 分子数: 3 / 由来タイプ: 組換発現 由来: (組換発現) ![]() 遺伝子: ptnD, LLKF_1870 / 発現宿主: ![]() ![]() 参照: UniProt: D2BKY9, protein-Npi-phosphohistidine-sugar phosphotransferase #3: タンパク質 | | 分子量: 11179.947 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() 遺伝子: lciA / 発現宿主: ![]() ![]() #4: タンパク質 | | 分子量: 5783.341 Da / 分子数: 1 / 由来タイプ: 組換発現 由来: (組換発現) ![]() 遺伝子: lcnA / 発現宿主: ![]() ![]() #5: 糖 | 研究の焦点であるリガンドがあるか | N | |
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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 |
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試料調製
構成要素 | 名称: The structure of LcnA, LciA, and the man-PTS of Lactococcus lactis タイプ: COMPLEX / Entity ID: #1-#4 / 由来: RECOMBINANT |
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由来(天然) | 生物種: ![]() |
由来(組換発現) | 生物種: ![]() ![]() |
緩衝液 | pH: 7 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
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 / 最大 デフォーカス(公称値): 1800 nm / 最小 デフォーカス(公称値): 1500 nm |
撮影 | 電子線照射量: 50 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) |
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解析
ソフトウェア | 名称: PHENIX / バージョン: 1.20.1_4487: / 分類: 精密化 | ||||||||||||||||||||||||
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CTF補正 | タイプ: NONE | ||||||||||||||||||||||||
3次元再構成 | 解像度: 2.98 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 1529899 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
原子モデル構築 | 空間: REAL | ||||||||||||||||||||||||
精密化 | 最高解像度: 2.98 Å | ||||||||||||||||||||||||
拘束条件 |
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