SASDCN3: Phosphoenolpyruvate-protein phosphotransferase (Enzyme I)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDCN3

試料

Phosphoenolpyruvate-protein phosphotransferase

• Phosphoenolpyruvate-protein phosphotransferase (protein), Enzyme I, Escherichia coli

機能・相同性

分子機能:

phosphoenolpyruvate-protein phosphotransferase / phosphoenolpyruvate-protein phosphotransferase activity / phosphoenolpyruvate-dependent sugar phosphotransferase system / kinase activity / metal ion binding / cytoplasm

ドメイン・相同性:

Phosphotransferase system, enzyme I / Phosphotransferase system, enzyme I-like / Phosphotransferase system, enzyme I N-terminal / PtsI, HPr-binding domain superfamily / PEP-utilising enzyme, N-terminal / PEP-utilising enzyme, active site / PEP-utilizing enzymes phosphorylation site signature. / PEP-utilising enzyme, conserved site / PEP-utilizing enzymes signature 2. / PEP-utilising enzyme, C-terminal / PEP-utilising enzyme, PEP-binding domain / PEP-utilising enzyme, mobile domain / Phosphohistidine domain superfamily / PEP-utilising enzyme, mobile domain / Pyruvate kinase-like domain superfamily / Pyruvate/Phosphoenolpyruvate kinase-like domain superfamily

構成要素:

Phosphoenolpyruvate-protein phosphotransferase

• 生物種: Escherichia coli (大腸菌)
• 引用: ジャーナル: J Am Chem Soc / 年: 2010; タイトル: Solution structure of the 128 kDa enzyme I dimer from Escherichia coli and its 146 kDa complex with HPr using residual dipolar couplings and small- and wide-angle X-ray scattering.; 著者: Charles D Schwieters / Jeong-Yong Suh / Alexander Grishaev / Rodolfo Ghirlando / Yuki Takayama / G Marius Clore / ; 要旨: The solution structures of free Enzyme I (EI, ∼128 kDa, 575 × 2 residues), the first enzyme in the bacterial phosphotransferase system, and its complex with HPr (∼146 kDa) have been solved using novel methodology that makes use of prior structural knowledge (namely, the structures of the dimeric EIC domain and the isolated EIN domain both free and complexed to HPr), combined with residual dipolar coupling (RDC), small- (SAXS) and wide- (WAXS) angle X-ray scattering and small-angle neutron scattering (SANS) data. The calculational strategy employs conjoined rigid body/torsion/Cartesian simulated annealing, and incorporates improvements in calculating and refining against SAXS/WAXS data that take into account complex molecular shapes in the description of the solvent layer resulting in a better representation of the SAXS/WAXS data. The RDC data orient the symmetrically related EIN domains relative to the C(2) symmetry axis of the EIC dimer, while translational, shape, and size information is provided by SAXS/WAXS. The resulting structures are independently validated by SANS. Comparison of the structures of the free EI and the EI-HPr complex with that of the crystal structure of a trapped phosphorylated EI intermediate reveals large (∼70-90°) hinge body rotations of the two subdomains comprising the EIN domain, as well as of the EIN domain relative to the dimeric EIC domain. These large-scale interdomain motions shed light on the structural transitions that accompany the catalytic cycle of EI.

モデル

• モデル #1248: ; タイプ: atomic / ソフトウェア: PDB / ダミー原子の半径: 1.90 A / カイ2乗値: 0.341; 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: Phosphoenolpyruvate-protein phosphotransferase / 試料濃度: 5 mg/ml
• バッファ: 名称: 20mM TRIS buffer, 100 mM NaCl, 10 mM DTT, 4 mM MgCl2, 1 mM EDTA; pH: 7.4 ; コメント: 1 tablet of protease inhibitor cocktail (SigmaFAST S8830) was added

要素 #658

名称: Enzyme I / タイプ: protein / 記述: Phosphoenolpyruvate-protein phosphotransferase / 分子量: 63.561 / 分子数: 2 / 由来: Escherichia coli / 参照: UniProt: A0A037YGN3
配列:

MISGILASPG IAFGKALLLK EDEIVIDRKK ISADQVDQEV ERFLSGRAKA SAQLETIKTK AGETFGEEKE AIFEGHIMLL EDEELEQEII ALIKDKHMTA DAAAHEVIEG QASALEELDD EYLKERAADV RDIGKRLLRN ILGLKIIDLS AIQDEVILVA ADLTPSETAQ LNLKKVLGFI TDAGGRTSHT SIMARSLELP AIVGTGSVTS QVKNDDYLIL DAVNNQVYVN PTNEVIDKMR AVQEQVASEK AELAKLKDLP AITLDGHQVE VCANIGTVRD VEGAERNGAE GVGLYRTEFL FMDRDALPTE EEQFAAYKAV AEACGSQAVI VRTMDIGGDK ELPYMNFPKE ENPFLGWRAI RIAMDRREIL RDQLRAILRA SAFGKLRIMF PMIISVEEVR ALRKEIEIYK QELRDEGKAF DESIEIGVMV ETPAAATIAR HLAKEVDFFS IGTNDLTQYT LAVDRGNDMI SHLYQPMSPS VLNLIKQVID ASHAEGKWTG MCGELAGDER ATLLLLGMGL DEFSMSAISI PRIKKIIRNT NFEDAKVLAE QALAQPTTDE LMTLVNKFIE EKTIC

実験情報

• ビーム: 設備名称: Advanced Photon Source (APS) 12ID-C / 地域: Argonne, IL / 国: USA / 線源: X-ray synchrotron / 波長: 0.06199 Å / スペクトロメータ・検出器間距離: 4 mm
• 検出器: 名称: Gold CCD

スキャン

タイトル: Phosphoenolpyruvate-protein phosphotransferase
測定日: 2010年8月23日 / セル温度: 25 °C / 照射時間: 0.25 sec. / フレーム数: 20 / 単位: 1/A /

	Min	Max
Q	0.0144	0.2208

距離分布関数 P(R)

ソフトウェア P(R): GNOM 5.0 / ポイント数: 164 /

	Min	Max
Q	0.014426	0.1953
P(R) point	1	164
R	0	148.6

結果

カーブのタイプ: single_conc /

	実験値	Porod
分子量	128 kDa	118 kDa
体積	-	189 nm3

	Guinier	P(R)	P(R) error	Guinier error
前方散乱 I0	0.0290498	-	-	-
慣性半径, Rg	4.1 nm	4.2 nm	0.02	0.06

	Max	Error
D	14.75	0.5