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データを開く
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基本情報
登録情報 | データベース: SASBDB / ID: SASDAL5 |
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![]() | CD27L
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生物種 | ![]() |
![]() | ![]() タイトル: The CD27L and CTP1L endolysins targeting Clostridia contain a built-in trigger and release factor. 著者: Matthew Dunne / Haydyn D T Mertens / Vasiliki Garefalaki / Cy M Jeffries / Andrew Thompson / Edward A Lemke / Dmitri I Svergun / Melinda J Mayer / Arjan Narbad / Rob Meijers / ![]() ![]() ![]() 要旨: The bacteriophage ΦCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, ...The bacteriophage ΦCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, and represents a promising alternative as a bactericide. To better understand the lysis mechanism, we have determined the crystal structure of an autoproteolytic fragment of the CD27L endolysin. The structure covers the C-terminal domain of the endolysin, and represents a novel fold that is identified in a number of lysins that target Clostridia bacteria. The structure indicates endolysin cleavage occurs at the stem of the linker connecting the catalytic domain with the C-terminal domain. We also solved the crystal structure of the C-terminal domain of a slow cleaving mutant of the CTP1L endolysin that targets C. tyrobutyricum. Two distinct dimerization modes are observed in the crystal structures for both endolysins, despite a sequence identity of only 22% between the domains. The dimers are validated to be present for the full length protein in solution by right angle light scattering, small angle X-ray scattering and cross-linking experiments using the cross-linking amino acid p-benzoyl-L-phenylalanine (pBpa). Mutagenesis on residues contributing to the dimer interfaces indicates that there is a link between the dimerization modes and the autocleavage mechanism. We show that for the CTP1L endolysin, there is a reduction in lysis efficiency that is proportional to the cleavage efficiency. We propose a model for endolysin triggering, where the extended dimer presents the inactive state, and a switch to the side-by-side dimer triggers the cleavage of the C-terminal domain. This leads to the release of the catalytic portion of the endolysin, enabling the efficient digestion of the bacterial cell wall. |
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構造の表示
構造ビューア | 分子: ![]() ![]() |
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ダウンロードとリンク
-モデル
モデル #269 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 1: head-head CD27L dimer / カイ2乗値: 2.4649 ![]() |
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モデル #270 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 2: side-side CD27L dimer / カイ2乗値: 2.4649 ![]() |
モデル #271 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 3a / カイ2乗値: 2.4649 ![]() |
モデル #272 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 3b / カイ2乗値: 2.4649 ![]() |
モデル #273 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / カイ2乗値: 2.4649 ![]() |
モデル #274 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 3c / カイ2乗値: 2.4649 ![]() |
モデル #275 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 3d / カイ2乗値: 2.4649 ![]() |
モデル #276 | ![]() タイプ: atomic / ダミー原子の半径: 1.90 A / コメント: OLIGOMER component 3e / カイ2乗値: 2.4649 ![]() |
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試料
![]() | 名称: CD27L / Sample MW: 64.2 kDa / 試料濃度: 0.90-4.00 / 濃度測定法: A280 |
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バッファ | 名称: 20 mM HEPES 150 mM NaCl / pH: 7.5 / 組成: 150 mM NaCl |
要素 #94 | 名称: CD27L / タイプ: protein / 記述: Clostridium difficile bacteriophage 27 endolysin / 分子量: 32.1 / 分子数: 2 / 由来: Clostridioides difficile 配列: MGSSHHHHHH SSGLVPRGSH MKICITVGHS ILKSGACTSA DGVVNEYQYN KSLAPVLADT FRKEGHKVDV IIsPEKQFKT KNEEKSYKIP RVNSGGYDLL IELHLNASNG QGKGSEVLYY SNKGLEYATR ICDKLGTVFK NRGAKLDKRL YILNSSKPTA VLIESFFCDN ...配列: MGSSHHHHHH SSGLVPRGSH MKICITVGHS ILKSGACTSA DGVVNEYQYN KSLAPVLADT FRKEGHKVDV IIsPEKQFKT KNEEKSYKIP RVNSGGYDLL IELHLNASNG QGKGSEVLYY SNKGLEYATR ICDKLGTVFK NRGAKLDKRL YILNSSKPTA VLIESFFCDN KEDYDKAKKL GHEGIAKLIV EGVLNKNINN EGVKQMYKHT IVYDGEVDKI SATVVGWGYN DGKILICDIK DYVPGQTQNL YVVGGGACEK ISSITKEKFI MIKGNDRFDT LYKALDFIN |
-実験情報
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検出器 | 名称: Pilatus 1M-W / Pixsize x: 0.172 mm | ||||||||||||||||||||||||||||||
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