+データを開く
-基本情報
登録情報 | データベース: SASBDB / ID: SASDE98 |
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試料 | Solution structure of Diadenylate cyclase (DacA) from Staphylococcus aureus
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機能・相同性 | 機能・相同性情報 : / diadenylate cyclase activity / diadenylate cyclase / cAMP biosynthetic process / adenylate cyclase activity / membrane => GO:0016020 / ATP binding / plasma membrane 類似検索 - 分子機能 |
生物種 | Staphylococcus aureus (strain USA300) (黄色ブドウ球菌) |
引用 | ジャーナル: PLoS Pathog / 年: 2019 タイトル: Inhibition of the Staphylococcus aureus c-di-AMP cyclase DacA by direct interaction with the phosphoglucosamine mutase GlmM. 著者: Tommaso Tosi / Fumiya Hoshiga / Charlotte Millership / Rahul Singh / Charles Eldrid / Delphine Patin / Dominique Mengin-Lecreulx / Konstantinos Thalassinos / Paul Freemont / Angelika Gründling / 要旨: c-di-AMP is an important second messenger molecule that plays a pivotal role in regulating fundamental cellular processes, including osmotic and cell wall homeostasis in many Gram-positive organisms. ...c-di-AMP is an important second messenger molecule that plays a pivotal role in regulating fundamental cellular processes, including osmotic and cell wall homeostasis in many Gram-positive organisms. In the opportunistic human pathogen Staphylococcus aureus, c-di-AMP is produced by the membrane-anchored DacA enzyme. Inactivation of this enzyme leads to a growth arrest under standard laboratory growth conditions and a re-sensitization of methicillin-resistant S. aureus (MRSA) strains to ß-lactam antibiotics. The gene coding for DacA is part of the conserved three-gene dacA/ybbR/glmM operon that also encodes the proposed DacA regulator YbbR and the essential phosphoglucosamine mutase GlmM, which is required for the production of glucosamine-1-phosphate, an early intermediate of peptidoglycan synthesis. These three proteins are thought to form a complex in vivo and, in this manner, help to fine-tune the cellular c-di-AMP levels. To further characterize this important regulatory complex, we conducted a comprehensive structural and functional analysis of the S. aureus DacA and GlmM enzymes by determining the structures of the S. aureus GlmM enzyme and the catalytic domain of DacA. Both proteins were found to be dimers in solution as well as in the crystal structures. Further site-directed mutagenesis, structural and enzymatic studies showed that multiple DacA dimers need to interact for enzymatic activity. We also show that DacA and GlmM form a stable complex in vitro and that S. aureus GlmM, but not Escherichia coli or Pseudomonas aeruginosa GlmM, acts as a strong inhibitor of DacA function without the requirement of any additional cellular factor. Based on Small Angle X-ray Scattering (SAXS) data, a model of the complex revealed that GlmM likely inhibits DacA by masking the active site of the cyclase and preventing higher oligomer formation. Together these results provide an important mechanistic insight into how c-di-AMP production can be regulated in the cell. |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-モデル
モデル #2648 | タイプ: dummy / ソフトウェア: (5.0) / ダミー原子の半径: 2.50 A / カイ2乗値: 1.140 Omokage検索でこの集合体の類似形状データを探す (詳細) |
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-試料
試料 | 名称: Solution structure of Diadenylate cyclase (DacA) from Staphylococcus aureus |
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バッファ | 名称: 30 mM Tris, 150 mM NaCl / pH: 7.5 |
要素 #1391 | 名称: DacA / タイプ: protein / 記述: Diadenylate cyclase / 分子量: 19.374 / 分子数: 2 / 由来: Staphylococcus aureus (strain USA300) / 参照: UniProt: A0A0H2XIU4 配列: GSHMASFLKR YTSNTYSKDE EKLIQSVSKA VQYMAKRRIG ALIVFEKETG LQDYIETGIA MDSNISQELL INVFIPNTPL HDGAMIIQGT KIAAAASYLP LSDSPKISKS LGTRHRAAVG ISEVSDAFTV IVSEETGDIS VTFDGKLRRD ISNEIFEELL AEHWFGTRFQ KKGVK |
-実験情報
ビーム | 設備名称: Diamond Light Source B21 / 地域: Oxfordshire / 国: UK / 形状: 1 x 5 mm / 線源: X-ray synchrotron / 波長: 0.1 Å / スペクトロメータ・検出器間距離: 4.014 mm | ||||||||||||||||||||||||||||||
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検出器 | 名称: Pilatus 2M | ||||||||||||||||||||||||||||||
スキャン | タイトル: Solution structure of Diadenylate cyclase (DacA) from Staphylococcus aureus 測定日: 2018年5月7日 / 保管温度: 20 °C / セル温度: 20 °C / 照射時間: 3 sec. / 単位: 1/A /
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距離分布関数 P(R) | ソフトウェア P(R): GNOM 5.0 / ポイント数: 1365 /
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結果 | カーブのタイプ: sec /
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