Q8X1D8 (NAO_FUSOX) Reviewed, UniProtKB/Swiss-Prot

Names and origin

Protein names	Recommended name: Nitroalkane oxidase Short name=NAO EC=1.7.3.1
Organism	Fusarium oxysporum (Panama disease fungus)
Taxonomic identifier	5507 [NCBI]
Taxonomic lineage	cellular organisms › Eukaryota › Opisthokonta › Fungi › Dikarya › Ascomycota › saccharomyceta › Pezizomycotina › leotiomyceta › sordariomyceta › Sordariomycetes › Hypocreomycetidae › Hypocreales › Nectriaceae › mitosporic Nectriaceae › Fusarium › Fusarium oxysporum species complex

Protein attributes

Sequence length	439 AA.
Sequence status	Complete.
Protein existence	Evidence at protein level

General annotation (Comments)

Function	Catalyzes the oxidative denitrification of neutral nitroalkanes, including 3-nitro-2-pentanol, 1-nitropropane, 2-nitropropane, nitroethane and nitrocyclohexane, and may thereby protect the organism against toxic compounds. Has no detectable acyl-CoA dehydrogenase activity. Ref.1 Ref.2 Ref.4
Catalytic activity	A nitroalkane + H2O + O2 = an aldehyde or ketone + nitrite + H2O2. Ref.1 Ref.2 Ref.3 Ref.5 Ref.6
Cofactor	FAD. Ref.1 Ref.2 Ref.4
Enzyme regulation	Strongly inhibited by mercury chloride and KCN. Ref.2
Subunit structure	Homotetramer. Ref.2 Ref.4
Induction	Up-regulated by nitroethane. Ref.2
Sequence similarities	Belongs to the acyl-CoA dehydrogenase family.
Biophysicochemical properties	Kinetic parameters: KM=1.54 mM for 1-nitropropane Ref.2 KM=7.4 mM for 2-nitropropane KM=1.0 mM for nitroethane KM=3.1 mM for 3-nitro-2-pentanol KM=0.9 mM for nitrocyclohexane KM=1.33 µM for FAD pH dependence: Optimum pH is 8.0. Temperature dependence: Optimum temperature is 40 degrees Celsius.

Ontologies

Keywords
Ligand	FAD Flavoprotein Nucleotide-binding
Molecular function	Oxidoreductase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological_process	nitrogen compound metabolic process Inferred from direct assay Ref.1. Source: UniProtKB
Molecular_function	FAD binding Inferred from direct assay Ref.1. Source: UniProtKB nitroalkane oxidase activity Inferred from direct assay Ref.1. Source: UniProtKB nitroethane oxidase activity Inferred from electronic annotation. Source: EC oxidoreductase activity, acting on the CH-CH group of donors Inferred from electronic annotation. Source: InterPro
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 439

439

Nitroalkane oxidase

PRO_0000418397

Regions

Nucleotide binding

131 – 134

4

FAD

Nucleotide binding

139 – 141

3

FAD

Nucleotide binding

169 – 171

3

FAD

Nucleotide binding

313 – 314

2

FAD

Nucleotide binding

375 – 379

5

FAD

Nucleotide binding

400 – 404

5

FAD

Sites

Active site

402

1

Proton acceptor Probable

Binding site

304

1

FAD

Experimental info

Mutagenesis

276

1

S → A: Decreases catalytic activity about tenfold. Ref.6

Mutagenesis

402

1

D → E: Decreases enzyme activity about twentyfold. Ref.1 Ref.3 Ref.6

Mutagenesis

402

1

D → N: Almost abolishes enzyme activity towards neutral nitroethane, but retains activity towards anionic nitroethane. Ref.1 Ref.3 Ref.6

Mutagenesis

409

1

R → K: Reduces catalytic activity. Ref.5

Secondary structure

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439

Helix Strand Turn

Details...

Helix

8 – 23

16

Helix

25 – 27

3

Helix

28 – 32

5

Helix

38 – 43

6

Helix

46 – 54

9

Turn

55 – 58

4

Helix

59 – 61

3

Helix

64 – 66

3

Helix

73 – 84

12

Helix

91 – 106

16

Helix

110 – 116

7

Helix

118 – 121

4

Beta strand

129 – 132

4

Turn

140 – 143

4

Beta strand

153 – 157

5

Beta strand

160 – 168

9

Turn

171 – 177

7

Beta strand

182 – 190

9

Helix

204 – 207

4

Beta strand

208 – 213

6

Helix

215 – 219

5

Helix

223 – 225

3

Beta strand

226 – 230

5

Beta strand

234 – 236

3

Beta strand

244 – 253

10

Helix

254 – 256

3

Helix

263 – 301

39

Beta strand

306 – 308

3

Helix

310 – 312

3

Helix

314 – 341

28

Beta strand

344 – 346

3

Helix

348 – 377

30

Helix

379 – 382

4

Helix

388 – 395

8

Turn

396 – 400

5

Beta strand

401 – 403

3

Turn

405 – 408

4

Helix

409 – 417

9

Helix

426 – 429

4

Sequences

	Sequence		Length	Mass (Da)
	Q8X1D8 [UniParc]. Last modified March 1, 2002. Version 1. Checksum: 9F156889F2A5901A	FASTA	439	48,162
	10 20 30 40 50 60 MVDFKLSPSQ LEARRHAQAF ANTVLTKASA EYSTQKDQLS RFQATRPFYR EAVRHGLIKA 70 80 90 100 110 120 QVPIPLGGTM ESLVHESIIL EELFAVEPAT SITIVATALG LMPVILCDSP SLQEKFLKPF 130 140 150 160 170 180 ISGEGEPLAS LMHSEPNGTA NWLQKGGPGL QTTARKVGNE WVISGEKLWP SNSGGWDYKG 190 200 210 220 230 240 ADLACVVCRV SDDPSKPQDP NVDPATQIAV LLVTRETIAN NKKDAYQILG EPELAGHITT 250 260 270 280 290 300 SGPHTRFTEF HVPHENLLCT PGLKAQGLVE TAFAMSAALV GAMAIGTARA AFEEALVFAK 310 320 330 340 350 360 SDTRGGSKHI IEHQSVADKL IDCKIRLETS RLLVWKAVTT LEDEALEWKV KLEMAMQTKI 370 380 390 400 410 420 YTTDVAVECV IDAMKAVGMK SYAKDMSFPR LLNEVMCYPL FDGGNIGLRR RQMQRVMALE 430 DYEPWAATYG SSKVDKSRL

References

[1]	"Cloning of nitroalkane oxidase from Fusarium oxysporum identifies a new member of the acyl-CoA dehydrogenase superfamily." Daubner S.C., Gadda G., Valley M.P., Fitzpatrick P.F. Proc. Natl. Acad. Sci. U.S.A. 99:2702-2707(2002) [PubMed] [Europe PMC] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [MRNA], PARTIAL PROTEIN SEQUENCE, CATALYTIC ACTIVITY, FUNCTION, MUTAGENESIS OF ASP-402, ACTIVE SITE, COFACTOR.
[2]	"Purification and properties of nitroalkane oxidase from Fusarium oxysporum." Kido T., Hashizume K., Soda K. J. Bacteriol. 133:53-58(1978) [PubMed] [Europe PMC] [Abstract] Cited for: CATALYTIC ACTIVITY, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, COFACTOR, ENZYME REGULATION, INDUCTION.
[3]	"Inactivation of nitroalkane oxidase upon mutation of the active site base and rescue with a deprotonated substrate." Valley M.P., Fitzpatrick P.F. J. Am. Chem. Soc. 125:8738-8739(2003) [PubMed] [Europe PMC] [Abstract] Cited for: CATALYTIC ACTIVITY, ACTIVE SITE, MUTAGENESIS OF ASP-402.
[4]	"Crystal structures of nitroalkane oxidase: insights into the reaction mechanism from a covalent complex of the flavoenzyme trapped during turnover." Nagpal A., Valley M.P., Fitzpatrick P.F., Orville A.M. Biochemistry 45:1138-1150(2006) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.07 ANGSTROMS) IN COMPLEX WITH NITROBUTANE AND FAD, FUNCTION, COFACTOR, ACTIVE SITE, SUBUNIT.
[5]	"Mechanistic and structural analyses of the roles of Arg409 and Asp402 in the reaction of the flavoprotein nitroalkane oxidase." Fitzpatrick P.F., Bozinovski D.M., Heroux A., Shaw P.G., Valley M.P., Orville A.M. Biochemistry 46:13800-13808(2007) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) OF MUTANT LYS-409 IN COMPLEX WITH FAD, CATALYTIC ACTIVITY, MUTAGENESIS OF ARG-409.
[6]	"Crystal structures of intermediates in the nitroalkane oxidase reaction." Heroux A., Bozinovski D.M., Valley M.P., Fitzpatrick P.F., Orville A.M. Biochemistry 48:3407-3416(2009) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF MUTANTS ALA-276 AND ASN-402 IN COMPLEXES WITH 1-NITROHEXANE AND FAD, CATALYTIC ACTIVITY, MUTAGENESIS OF SER-276 AND ASP-402.
[7]	"Differential quantum tunneling contributions in nitroalkane oxidase catalyzed and the uncatalyzed proton transfer reaction." Major D.T., Heroux A., Orville A.M., Valley M.P., Fitzpatrick P.F., Gao J. Proc. Natl. Acad. Sci. U.S.A. 106:20734-20739(2009) [PubMed] [Europe PMC] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) OF MUTANT ASN-402 IN COMPLEX WITH FAD AND NITROETHANE.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ

AF425595 mRNA. Translation: AAL57485.1.

3D structure databases

PDBe
RCSB PDB
PDBj

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
2C0U	X-ray	2.20	A/B/C/D	1-439	[»]
2C12	X-ray	2.07	A/B/C/D/E/F	1-439	[»]
2REH	X-ray	2.40	A/B/C/D	1-439	[»]
2ZAF	X-ray	2.50	A/B/C/D	1-439	[»]
3D9D	X-ray	2.10	A/B/C/D	2-439	[»]
3D9E	X-ray	2.20	A/B/C/D	2-439	[»]
3D9F	X-ray	2.20	A/B/C/D	2-439	[»]
3D9G	X-ray	2.15	A/B/C/D	2-439	[»]
3FCJ	X-ray	2.40	A/B/C/D	2-439	[»]

ProteinModelPortal

Q8X1D8.

SMR

Q8X1D8. Positions 2-431.

ModBase

Search...

Protocols and materials databases

StructuralBiologyKnowledgebase

Search...

Enzyme and pathway databases

BioCyc

MetaCyc:MONOMER-12547.

SABIO-RK

Q8X1D8.

Family and domain databases

Gene3D

1.10.540.10. 1 hit.
2.40.110.10. 1 hit.

InterPro

IPR006092. Acyl-CoA_DH_N.
IPR006090. Acyl-CoA_Oxase/DH_1.
IPR006091. Acyl-CoA_Oxase/DH_cen-dom.
IPR009075. AcylCo_DH/oxidase_C.
IPR013786. AcylCoA_DH/ox_N.
IPR009100. AcylCoA_DH/oxidase.
[Graphical view]

Pfam

PF00441. Acyl-CoA_dh_1. 1 hit.
PF02770. Acyl-CoA_dh_M. 1 hit.
PF02771. Acyl-CoA_dh_N. 1 hit.
[Graphical view]

SUPFAM

SSF56645. AcylCoA_dehyd_NM. 1 hit.
SSF47203. AcylCoADH_C_like. 1 hit.

PROSITE

PS00072. ACYL_COA_DH_1. False negative.
PS00073. ACYL_COA_DH_2. False negative.
[Graphical view]

ProtoNet

Search...

Other

EvolutionaryTrace

Q8X1D8.

Entry information

Entry name

NAO_FUSOX

Accession

Primary (citable) accession number: Q8X1D8

Entry history

Integrated into UniProtKB/Swiss-Prot:	September 5, 2012
Last sequence update:	March 1, 2002
Last modified:	April 3, 2013
This is version 70 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation program

Fungal Protein Annotation Program