SASDA45: PpoA wild type enzyme (Psi-producing oxygenase A, PpoA)

基本情報

• 登録情報: データベース: SASBDB / ID: SASDA45

試料

PpoA wild type enzyme

• Psi-producing oxygenase A (protein), PpoA, Aspergillus nidulans

機能・相同性

分子機能:

linoleate 8R-lipoxygenase / 9,12-octadecadienoate 8-hydroperoxide 8R-isomerase / linoleate 8R-lipoxygenase activity / oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen / isomerase activity / peroxidase activity / fatty acid metabolic process / monooxygenase activity / response to oxidative stress / iron ion binding / heme binding

ドメイン・相同性:

Psi-factor producing oxygenase, N-terminal heme peroxidase domain / : / Haem peroxidase, animal-type / Haem peroxidase domain superfamily, animal type / Animal haem peroxidase / Animal heme peroxidase superfamily profile. / Haem peroxidase superfamily / Cytochrome P450, conserved site / Cytochrome P450 cysteine heme-iron ligand signature. / Cytochrome P450 / Cytochrome P450 superfamily / Cytochrome P450

構成要素:

Psi-producing oxygenase A

• 生物種: Aspergillus nidulans (カビ)
• 引用: ジャーナル: Biochim Biophys Acta / 年: 2013; タイトル: A structural model of PpoA derived from SAXS-analysis-implications for substrate conversion.; 著者: Christian Koch / Giancarlo Tria / Alistair J Fielding / Florian Brodhun / Oliver Valerius / Kirstin Feussner / Gerhard H Braus / Dmitri I Svergun / Marina Bennati / Ivo Feussner / ; 要旨: In plants and mammals, oxylipins may be synthesized via multi step processes that consist of dioxygenation and isomerization of the intermediately formed hydroperoxy fatty acid. These processes are typically catalyzed by two distinct enzyme classes: dioxygenases and cytochrome P450 enzymes. In ascomycetes biosynthesis of oxylipins may proceed by a similar two-step pathway. An important difference, however, is that both enzymatic activities may be combined in a single bifunctional enzyme. These types of enzymes are named Psi-factor producing oxygenases (Ppo). Here, the spatial organization of the two domains of PpoA from Aspergillus nidulans was analyzed by small-angle X-ray scattering and the obtained data show that the enzyme exhibits a relatively flat trimeric shape. Atomic structures of the single domains were obtained by template-based structure prediction and docked into the enzyme envelope of the low resolution structure obtained by SAXS. EPR-based distance measurements between the tyrosyl radicals formed in the activated dioxygenase domain of the enzyme supported the trimeric structure obtained from SAXS and the previous assignment of Tyr374 as radical-site in PpoA. Furthermore, two phenylalanine residues in the cytochrome P450 domain were shown to modulate the specificity of hydroperoxy fatty acid rearrangement.

登録者

• Giancarlo Tria (EMBL-Hamburg, European Molecular Biology Laboratory (EMBL) - Hamburg outstation, Notkestraße 85, Geb. 25A, 22607 Hamburg, Deutschland, Germany)

モデル

• モデル #93: ; タイプ: atomic / カイ2乗値: 0.4761; 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: PpoA wild type enzyme / Sample MW: 362.34 kDa / 試料濃度: 0.40-26.00
• バッファ: 名称: 20 Mm HEPES / 濃度: 20.00 mM / pH: 7.5

要素 #77

名称: PpoA / タイプ: protein / 記述: Psi-producing oxygenase A / 分子量: 120.78 / 分子数: 3 / 由来: Aspergillus nidulans / 参照: UniProt: Q6RET3
配列:

MGEDKETNIL AGLGNTISQV ENVVAASLRP LPTATGDGTY VAESTQTGLA KDLSHVDLKD VRTLAEVVKS AATGEPVDDK QYIMERVIQL AAGLPSTSRN AAELTKSFLN MLWNDLEHPP VSYLGADSMH RKADGSGNNR FWPQLGAAGS AYARSVRPKT MQSPSLPDPE TIFDCLLRRK EYREHPNKIS SVLFYLASII IHDLFQTDPK DNSVSKTSSY LDLSPLYGNN QDEQNLVRTF KDGKLKPDCF ATKRVLGFPP GVGVLLIMFN RFHNYVVDQL AAINECGRFT KPDESNVDEY AKYDNNLFQT GRLVTCGLYA NIILKDYVRT ILNINRTDST WSLDPRMEMK DGLLGEAAAM ATGNQVSAEF NVVYRWHACI SKRDEKWTED FHREIMPGVD PSTLSMQDFV AGLGRWQAGL PQEPLERPFS GLQRKPDGAF NDDDLVNLFE KSVEDCAGAF GASHVPAIFK SVEALGIMQA RRWNLGTLNE FRQYFNLAPH KTFEDINSDP YIADQLKRLY DHPDLVEIYP GVVVEEAKDS MVPGSGLCTN FTISRAILSD AVALVRGDRF YTVDYTPKHL TNWAYNEIQP NNAVDQGQVF YKLVLRAFPN HFDGNSIYAH FPLVVPSENE KILKSLGVAE KYSWEKPSRI SHPIFISSHA ACMSILENQE TFKVTWGRKI EFLMQRDKHQ YGKDFMLSGD RPPNAASRKM MGSALYRDEW EAEVKNFYEQ TTLKLLHKNS YKLAGVNQVD IVRDVANLAQ VHFCSSVFSL PLKTDSNPRG IFAESELYKI MAAVFTAIFY DADIGKSFEL NQAARTVTQQ LGQLTMANVE IIAKTGLIAN LVNRLHRRDV LSEYGIHMIQ RLLDSGLPAT EIVWTHILPT AGGMVANQAQ LFSQCLDYYL SEEGSGHLPE INRLAKENTP EADELLTRYF MEGARLRSSV ALPRVAAQPT VVEDNGEKLT IKAGQVVMCN LVSACMDPTA FPDPEKVKLD RDMNLYAHFG FGPHKCLGLD LCKTGLSTML KVLGRLDNLR RAPGAQGQLK KLSGPGGIAK YMNEDQSGFT PFPSTMKIQW DGELPQLKED F

実験情報

• ビーム: 設備名称: DORIS III X33 / 地域: Hamburg / 国: Germany / 形状: 0.6 / 線源: X-ray synchrotron / 波長: 0.15 Å / スペクトロメータ・検出器間距離: 2.7 mm
• 検出器: 名称: Pilatus 1M-W / Pixsize x: 0.172 mm

スキャン

タイトル: PpoA wild type enzyme / 測定日: 2011年6月24日 / 保管温度: 10 °C / セル温度: 10 °C / 照射時間: 120 sec. / フレーム数: 8 / 単位: 1/nm /

	Min	Max
Q	0.0855	6.0292

距離分布関数 P(R)

ソフトウェア P(R): GNOM 4.6 / ポイント数: 290 /

	Min	Max
Q	0.1115	1.697
P(R) point	10	299
R	0	16.5

結果

カーブのタイプ: single_conc /

	実験値	Porod	Estimated
分子量	330 kDa	-	-
体積	-	540 nm3	667

	Guinier	P(R)
前方散乱 I0	26.8	-
慣性半径, Rg	5.4 nm	5.4 nm

	Min	Max
D	-	16.5
Guinier point	14	44