SASDCL9: Catalytic domain (AC) of B. Pertussis Adenylate Cyclase Toxin (Cy...

基本情報

• 登録情報: データベース: SASBDB / ID: SASDCL9

試料

Catalytic domain (AC) of B. Pertussis Adenylate Cyclase Toxin (CyaA) in complex with calmodulin

• Bifunctional hemolysin/adenylate cyclase (protein), AC domain from CyaA, Bordetella pertussis (strain Tohama I / ATCC BAA-589 / NCTC 13251)
• Calmodulin (protein), CaM, Homo sapiens

機能・相同性

分子機能:

calcium- and calmodulin-responsive adenylate cyclase activity / hemolysis in another organism / : / adenylate cyclase / establishment of protein localization to mitochondrial membrane / type 3 metabotropic glutamate receptor binding / cAMP biosynthetic process / adenylate cyclase activity / channel activity / regulation of synaptic vesicle endocytosis / negative regulation of high voltage-gated calcium channel activity / positive regulation of cyclic-nucleotide phosphodiesterase activity / negative regulation of calcium ion export across plasma membrane / regulation of synaptic vesicle exocytosis / regulation of cardiac muscle cell action potential / response to corticosterone / nitric-oxide synthase binding / positive regulation of ryanodine-sensitive calcium-release channel activity / regulation of cell communication by electrical coupling involved in cardiac conduction / negative regulation of peptidyl-threonine phosphorylation / protein phosphatase activator activity / positive regulation of phosphoprotein phosphatase activity / adenylate cyclase binding / catalytic complex / detection of calcium ion / regulation of cardiac muscle contraction / negative regulation of ryanodine-sensitive calcium-release channel activity / positive regulation of DNA binding / regulation of cardiac muscle contraction by regulation of the release of sequestered calcium ion / phosphatidylinositol 3-kinase binding / regulation of release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / enzyme regulator activity / positive regulation of protein dephosphorylation / regulation of calcium-mediated signaling / titin binding / positive regulation of protein autophosphorylation / voltage-gated potassium channel complex / calcium channel complex / response to amphetamine / activation of adenylate cyclase activity / substantia nigra development / adenylate cyclase activator activity / nitric-oxide synthase regulator activity / regulation of heart rate / sarcomere / protein serine/threonine kinase activator activity / regulation of cytokinesis / positive regulation of peptidyl-threonine phosphorylation / positive regulation of nitric-oxide synthase activity / mitochondrial membrane / positive regulation of protein serine/threonine kinase activity / spindle microtubule / synaptic vesicle membrane / spindle pole / response to calcium ion / G2/M transition of mitotic cell cycle / calcium-dependent protein binding / disordered domain specific binding / myelin sheath / toxin activity / positive regulation of cytosolic calcium ion concentration / growth cone / vesicle / transmembrane transporter binding / calmodulin binding / G protein-coupled receptor signaling pathway / protein domain specific binding / centrosome / calcium ion binding / protein kinase binding / host cell plasma membrane / protein-containing complex / extracellular region / ATP binding / membrane / nucleus / plasma membrane / cytoplasm

ドメイン・相同性:

RTX, pore-forming domain / N-terminal domain in RTX protein / Haemolysin-type calcium binding-related / Haemolysin-type calcium binding protein related domain / RTX toxin determinant A / Anthrax toxin, edema factor, central / Anthrax toxin, edema factor, C-terminal / Anthrax toxin, edema factor, central domain superfamily / Anthrax toxin LF subunit / Hemolysin-type calcium-binding conserved site / Hemolysin-type calcium-binding region signature. / RTX calcium-binding nonapeptide repeat / RTX calcium-binding nonapeptide repeat (4 copies) / Serralysin-like metalloprotease, C-terminal / EF-hand domain pair / EF-hand, calcium binding motif / EF-Hand 1, calcium-binding site / EF-hand calcium-binding domain. / EF-hand calcium-binding domain profile. / EF-hand domain / EF-hand domain pair

構成要素:

Bifunctional hemolysin/adenylate cyclase / Calmodulin-3

• 生物種: Bordetella pertussis (strain Tohama I / ATCC BAA-589 / NCTC 13251) (百日咳菌); Homo sapiens (ヒト)
• 引用: ジャーナル: PLoS Biol / 年: 2017; タイトル: Calmodulin fishing with a structurally disordered bait triggers CyaA catalysis.; 著者: Darragh P O'Brien / Dominique Durand / Alexis Voegele / Véronique Hourdel / Marilyne Davi / Julia Chamot-Rooke / Patrice Vachette / Sébastien Brier / Daniel Ladant / Alexandre Chenal / ; 要旨: Once translocated into the cytosol of target cells, the catalytic domain (AC) of the adenylate cyclase toxin (CyaA), a major virulence factor of Bordetella pertussis, is potently activated by binding calmodulin (CaM) to produce supraphysiological levels of cAMP, inducing cell death. Using a combination of small-angle X-ray scattering (SAXS), hydrogen/deuterium exchange mass spectrometry (HDX-MS), and synchrotron radiation circular dichroism (SR-CD), we show that, in the absence of CaM, AC exhibits significant structural disorder, and a 75-residue-long stretch within AC undergoes a disorder-to-order transition upon CaM binding. Beyond this local folding, CaM binding induces long-range allosteric effects that stabilize the distant catalytic site, whilst preserving catalytic loop flexibility. We propose that the high enzymatic activity of AC is due to a tight balance between the CaM-induced decrease of structural flexibility around the catalytic site and the preservation of catalytic loop flexibility, allowing for fast substrate binding and product release. The CaM-induced dampening of AC conformational disorder is likely relevant to other CaM-activated enzymes.

登録者

• Dominique DURAND (I2BC-CNRS)

モデル

• モデル #1694: ; タイプ: mix / ソフトウェア: (08) / ダミー原子の半径: 1.90 A / カイ2乗値: 1.956; Omokage検索でこの集合体の類似形状データを探す (詳細)
• モデル #1696: ; タイプ: atomic / ソフトウェア: (2.1) / ダミー原子の半径: 1.90 A / カイ2乗値: 1.410 / P-value: 0.000713; Omokage検索でこの集合体の類似形状データを探す (詳細)

試料

• 試料: 名称: Catalytic domain (AC) of B. Pertussis Adenylate Cyclase Toxin (CyaA) in complex with calmodulin; Entity id: 904 / 905
• バッファ: 名称: 20 mM Hepes, 150 mM NaCl, 4 mM CaCl2 / pH: 7.4

要素 #904

名称: AC domain from CyaA / タイプ: protein / 記述: Bifunctional hemolysin/adenylate cyclase / 分子量: 39.38 / 分子数: 1
由来: Bordetella pertussis (strain Tohama I / ATCC BAA-589 / NCTC 13251)
参照: UniProt: P0DKX7
配列:

MQQSHQAGYA NAADRESGIP AAVLDGIKAV AKEKNATLMF RLVNPHSTSL IAEGVATKGL GVHAKSSDWG LQAGYIPVNP NLSKLFGRAP EVIARADNDV NSSLAHGHTA VDLTLSKERL DYLRQAGLVT GMADGVVASN HAGYEQFEFR VKETSDGRYA VQYRRKGGDD FEAVKVIGNA AGIPLTADID MFAIMPHLSN FRDSARSSVT SGDSVTDYLA RTRRAASEAT GGLDRERIDL LWKIARAGAR SAVGTEARRQ FRYDGDMNIG VITDFELEVR NALNRRAHAV GAQDVVQHGT EQNNPFPEAD EKIFVVSATG ESQMLTRGQL KEYIGQQRGE GYVFYENRAY GVAGKSLFDD GLGA

要素 #905

名称: CaM / タイプ: protein / 記述: Calmodulin / 分子量: 16.837 / 分子数: 1 / 由来: Homo sapiens / 参照: UniProt: P62158
配列:

MADQLTEEQI AEFKEAFSLF DKDGDGTITT KELGTVMRSL GQNPTEAELQ DMINEVDADG NGTIDFPEFL TMMARKMKDT DSEEEIREAF RVFDKDGNGY ISAAELRHVM TNLGEKLTDE EVDEMIREAD IDGDGQVNYE EFVQMMTAK

実験情報

• ビーム: 設備名称: SOLEIL SWING / 地域: Saint-Aubin / 国: France / 線源: X-ray synchrotron / 波長: 0.1 Å / スペクトロメータ・検出器間距離: 1.987 mm
• 検出器: 名称: AVIEX / タイプ: CCD

スキャン

タイトル: Catalytic domain (AC) of B. Pertussis Adenylate Cyclase Toxin (CyaA) in complex with calmodulin
測定日: 2015年6月19日 / 保管温度: 10 °C / セル温度: 15 °C / 照射時間: 1.5 sec. / フレーム数: 250 / 単位: 1/A /

	Min	Max
Q	0.0061	0.4004

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 775 /

	Min	Max
Q	0.00612344	0.400433
P(R) point	1	775
R	0	98

結果

カーブのタイプ: sec
コメント: The scattered intensities were displayed on an absolute scale (cm-1) using the scattering of water. Frames were examined individually and 20 identical frames were averaged and further processed. The corresponding concentration was 0.82 g/L. Three independent determinations of the molecular mass were obtained from the value of I(0)/c, where c is the protein concentration, and using the programs SAXSMow2 and ScÅtter3 available at the URLs http://saxs.ifsc.usp.br/ and https://bl1231.als.lbl.gov/scatter/, respectively. The average value is The average value is MWexperimental=56.3 kDa. AC-CAM complex: Top panel: Comparison of the experimental data (blue dots) with the calculated scattering pattern (red line) of the BUNCH model shown on the right. chi2=1.96. Each CaM domain were handled as rigid bodies while the program searches an optimal conformation of the inter-domain helix of CaM. Bottom panel: Typical ensemble of conformations describing the AC:CaM complex, obtained using the program EOM and displayed after superimposition of the AC moiety of each conformation. chi2=1.41. The corresponding scattering curve is shown in red superimposed over experimental data (blue dots).

	実験値	Standard	Standard error	Porod
分子量	56.3 kDa	56.3 kDa	3	51.9 kDa
体積	-	-	-	77.9 nm3

	P(R)	P(R) error	Guinier	Guinier error
前方散乱 I0	0.0383	0.0003	0.0383	0.0003
慣性半径, Rg	2.97 nm	0.03	2.92 nm	0.03

	Min	Max	Error
D	-	9.8	0.5
Guinier point	1	58	-