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

Basic information

• Entry: Database: SASBDB / ID: SASDA45

Sample

PpoA wild type enzyme

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

Function / homology

Function:

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

Domain/homology:

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

Component:

Psi-producing oxygenase A

• Biological species: Aspergillus nidulans (mold)
• Citation: Journal: Biochim Biophys Acta / Year: 2013; Title: A structural model of PpoA derived from SAXS-analysis-implications for substrate conversion.; Authors: Christian Koch / Giancarlo Tria / Alistair J Fielding / Florian Brodhun / Oliver Valerius / Kirstin Feussner / Gerhard H Braus / Dmitri I Svergun / Marina Bennati / Ivo Feussner / ; Abstract: 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.

Contact author

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

Models

• Model #93: ; Type: atomic / Chi-square value: 0.4761; Search similar-shape structures of this assembly by Omokage search (details)

Sample

• Sample: Name: PpoA wild type enzyme / Sample MW: 362.34 kDa / Specimen concentration: 0.40-26.00
• Buffer: Name: 20 Mm HEPES / Concentration: 20.00 mM / pH: 7.5

Entity #77

Name: PpoA / Type: protein / Description: Psi-producing oxygenase A / Formula weight: 120.78 / Num. of mol.: 3 / Source: Aspergillus nidulans / References: UniProt: Q6RET3
Sequence:

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

Experimental information

• Beam: Instrument name: DORIS III X33 / City: Hamburg / 国: Germany / Shape: 0.6 / Type of source: X-ray synchrotron / Wavelength: 0.15 Å / Dist. spec. to detc.: 2.7 mm
• Detector: Name: Pilatus 1M-W / Pixsize x: 0.172 mm

Scan

Title: PpoA wild type enzyme / Measurement date: Jun 24, 2011 / Storage temperature: 10 °C / Cell temperature: 10 °C / Exposure time: 120 sec. / Number of frames: 8 / Unit: 1/nm /

	Min	Max
Q	0.0855	6.0292

Distance distribution function P(R)

Sofotware P(R): GNOM 4.6 / Number of points: 290 /

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

Result

Type of curve: single_conc /

	Experimental	Porod	Estimated
MW	330 kDa	-	-
Volume	-	540 nm3	667

	Guinier	P(R)
Forward scattering, I0	26.8	-
Radius of gyration, Rg	5.4 nm	5.4 nm

	Min	Max
D	-	16.5
Guinier point	14	44