NAO_FUSOX
ID NAO_FUSOX Reviewed; 439 AA.
AC Q8X1D8;
DT 05-SEP-2012, integrated into UniProtKB/Swiss-Prot.
DT 01-MAR-2002, sequence version 1.
DT 03-AUG-2022, entry version 101.
DE RecName: Full=Nitroalkane oxidase;
DE Short=NAO;
DE EC=1.7.3.1;
OS Fusarium oxysporum (Fusarium vascular wilt).
OC Eukaryota; Fungi; Dikarya; Ascomycota; Pezizomycotina; Sordariomycetes;
OC Hypocreomycetidae; Hypocreales; Nectriaceae; Fusarium;
OC Fusarium oxysporum species complex.
OX NCBI_TaxID=5507;
RN [1]
RP NUCLEOTIDE SEQUENCE [MRNA], PARTIAL PROTEIN SEQUENCE, CATALYTIC ACTIVITY,
RP FUNCTION, MUTAGENESIS OF ASP-402, ACTIVE SITE, AND COFACTOR.
RC STRAIN=ATCC 695;
RX PubMed=11867731; DOI=10.1073/pnas.052527799;
RA Daubner S.C., Gadda G., Valley M.P., Fitzpatrick P.F.;
RT "Cloning of nitroalkane oxidase from Fusarium oxysporum identifies a new
RT member of the acyl-CoA dehydrogenase superfamily.";
RL Proc. Natl. Acad. Sci. U.S.A. 99:2702-2707(2002).
RN [2]
RP CATALYTIC ACTIVITY, FUNCTION, BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT,
RP COFACTOR, ACTIVITY REGULATION, AND INDUCTION.
RC STRAIN=ATCC 659 / NBRC 5942 / DR 1004;
RX PubMed=22538; DOI=10.1128/jb.133.1.53-58.1978;
RA Kido T., Hashizume K., Soda K.;
RT "Purification and properties of nitroalkane oxidase from Fusarium
RT oxysporum.";
RL J. Bacteriol. 133:53-58(1978).
RN [3]
RP CATALYTIC ACTIVITY, ACTIVE SITE, AND MUTAGENESIS OF ASP-402.
RX PubMed=12862464; DOI=10.1021/ja036045s;
RA Valley M.P., Fitzpatrick P.F.;
RT "Inactivation of nitroalkane oxidase upon mutation of the active site base
RT and rescue with a deprotonated substrate.";
RL J. Am. Chem. Soc. 125:8738-8739(2003).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.07 ANGSTROMS) IN COMPLEX WITH THE SUBSTRATE
RP NITROBUTANE AND FAD, FUNCTION, COFACTOR, ACTIVE SITE, AND SUBUNIT.
RX PubMed=16430210; DOI=10.1021/bi051966w;
RA Nagpal A., Valley M.P., Fitzpatrick P.F., Orville A.M.;
RT "Crystal structures of nitroalkane oxidase: insights into the reaction
RT mechanism from a covalent complex of the flavoenzyme trapped during
RT turnover.";
RL Biochemistry 45:1138-1150(2006).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) OF MUTANT LYS-409 IN COMPLEX WITH
RP FAD, COFACTOR, CATALYTIC ACTIVITY, AND MUTAGENESIS OF ARG-409.
RX PubMed=17994768; DOI=10.1021/bi701557k;
RA Fitzpatrick P.F., Bozinovski D.M., Heroux A., Shaw P.G., Valley M.P.,
RA Orville A.M.;
RT "Mechanistic and structural analyses of the roles of Arg409 and Asp402 in
RT the reaction of the flavoprotein nitroalkane oxidase.";
RL Biochemistry 46:13800-13808(2007).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.10 ANGSTROMS) OF MUTANTS ALA-276 AND ASN-402 IN
RP COMPLEXES WITH THE SUBSTRATE 1-NITROHEXANE AND FAD, CATALYTIC ACTIVITY,
RP COFACTOR, AND MUTAGENESIS OF SER-276 AND ASP-402.
RX PubMed=19265437; DOI=10.1021/bi8023042;
RA Heroux A., Bozinovski D.M., Valley M.P., Fitzpatrick P.F., Orville A.M.;
RT "Crystal structures of intermediates in the nitroalkane oxidase reaction.";
RL Biochemistry 48:3407-3416(2009).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) OF MUTANT ASN-402 IN COMPLEX WITH
RP FAD AND THE SUBSTRATE NITROETHANE, AND COFACTOR.
RX PubMed=19926855; DOI=10.1073/pnas.0911416106;
RA Major D.T., Heroux A., Orville A.M., Valley M.P., Fitzpatrick P.F., Gao J.;
RT "Differential quantum tunneling contributions in nitroalkane oxidase
RT catalyzed and the uncatalyzed proton transfer reaction.";
RL Proc. Natl. Acad. Sci. U.S.A. 106:20734-20739(2009).
CC -!- FUNCTION: Catalyzes the oxidative denitrification of neutral
CC nitroalkanes, including 3-nitro-2-pentanol, 1-nitropropane, 2-
CC nitropropane, nitroethane and nitrocyclohexane, and may thereby protect
CC the organism against toxic compounds. Has no detectable acyl-CoA
CC dehydrogenase activity. {ECO:0000269|PubMed:11867731,
CC ECO:0000269|PubMed:16430210, ECO:0000269|PubMed:22538}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a primary nitroalkane + H2O + O2 = an aldehyde + H(+) + H2O2 +
CC nitrite; Xref=Rhea:RHEA:20976, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16240, ChEBI:CHEBI:16301,
CC ChEBI:CHEBI:17478, ChEBI:CHEBI:133972; EC=1.7.3.1;
CC Evidence={ECO:0000269|PubMed:11867731, ECO:0000269|PubMed:12862464,
CC ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19265437,
CC ECO:0000269|PubMed:22538};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=a secondary nitroalkane + H2O + O2 = a ketone + H(+) + H2O2 +
CC nitrite; Xref=Rhea:RHEA:26490, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16240, ChEBI:CHEBI:16301,
CC ChEBI:CHEBI:17087, ChEBI:CHEBI:139218; EC=1.7.3.1;
CC Evidence={ECO:0000269|PubMed:11867731, ECO:0000269|PubMed:12862464,
CC ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19265437,
CC ECO:0000269|PubMed:22538};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:11867731, ECO:0000269|PubMed:16430210,
CC ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19265437,
CC ECO:0000269|PubMed:19926855, ECO:0000269|PubMed:22538};
CC -!- ACTIVITY REGULATION: Strongly inhibited by mercury chloride and KCN.
CC {ECO:0000269|PubMed:22538}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=1.54 mM for 1-nitropropane {ECO:0000269|PubMed:22538};
CC KM=7.4 mM for 2-nitropropane {ECO:0000269|PubMed:22538};
CC KM=1.0 mM for nitroethane {ECO:0000269|PubMed:22538};
CC KM=3.1 mM for 3-nitro-2-pentanol {ECO:0000269|PubMed:22538};
CC KM=0.9 mM for nitrocyclohexane {ECO:0000269|PubMed:22538};
CC KM=1.33 uM for FAD {ECO:0000269|PubMed:22538};
CC pH dependence:
CC Optimum pH is 8.0. {ECO:0000269|PubMed:22538};
CC Temperature dependence:
CC Optimum temperature is 40 degrees Celsius.
CC {ECO:0000269|PubMed:22538};
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:16430210,
CC ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855,
CC ECO:0000269|PubMed:22538}.
CC -!- INDUCTION: Up-regulated by nitroethane. {ECO:0000269|PubMed:22538}.
CC -!- SIMILARITY: Belongs to the acyl-CoA dehydrogenase family.
CC {ECO:0000305}.
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DR EMBL; AF425595; AAL57485.1; -; mRNA.
DR PDB; 2C0U; X-ray; 2.20 A; A/B/C/D=1-439.
DR PDB; 2C12; X-ray; 2.07 A; A/B/C/D/E/F=1-439.
DR PDB; 2REH; X-ray; 2.40 A; A/B/C/D=1-439.
DR PDB; 2ZAF; X-ray; 2.50 A; A/B/C/D=1-439.
DR PDB; 3D9D; X-ray; 2.10 A; A/B/C/D=2-439.
DR PDB; 3D9E; X-ray; 2.20 A; A/B/C/D=2-439.
DR PDB; 3D9F; X-ray; 2.20 A; A/B/C/D=2-439.
DR PDB; 3D9G; X-ray; 2.15 A; A/B/C/D=2-439.
DR PDB; 3FCJ; X-ray; 2.40 A; A/B/C/D=2-439.
DR PDBsum; 2C0U; -.
DR PDBsum; 2C12; -.
DR PDBsum; 2REH; -.
DR PDBsum; 2ZAF; -.
DR PDBsum; 3D9D; -.
DR PDBsum; 3D9E; -.
DR PDBsum; 3D9F; -.
DR PDBsum; 3D9G; -.
DR PDBsum; 3FCJ; -.
DR AlphaFoldDB; Q8X1D8; -.
DR SMR; Q8X1D8; -.
DR KEGG; ag:AAL57485; -.
DR VEuPathDB; FungiDB:FOC1_g10009155; -.
DR VEuPathDB; FungiDB:FOC4_g10012108; -.
DR VEuPathDB; FungiDB:FOIG_12289; -.
DR VEuPathDB; FungiDB:FOMG_12667; -.
DR VEuPathDB; FungiDB:FOXG_08703; -.
DR VEuPathDB; FungiDB:FOZG_06512; -.
DR VEuPathDB; FungiDB:HZS61_013072; -.
DR BioCyc; MetaCyc:MON-12547; -.
DR BRENDA; 1.7.3.1; 2351.
DR SABIO-RK; Q8X1D8; -.
DR EvolutionaryTrace; Q8X1D8; -.
DR GO; GO:0071949; F:FAD binding; IDA:UniProtKB.
DR GO; GO:0052664; F:nitroalkane oxidase activity; IDA:UniProtKB.
DR GO; GO:0050141; F:nitroethane oxidase activity; IEA:UniProtKB-EC.
DR GO; GO:0016627; F:oxidoreductase activity, acting on the CH-CH group of donors; IEA:InterPro.
DR GO; GO:0006807; P:nitrogen compound metabolic process; IDA:UniProtKB.
DR Gene3D; 1.10.540.10; -; 1.
DR Gene3D; 2.40.110.10; -; 1.
DR InterPro; IPR046373; Acyl-CoA_Oxase/DH_mid-dom_sf.
DR InterPro; IPR036250; AcylCo_DH-like_C.
DR InterPro; IPR009075; AcylCo_DH/oxidase_C.
DR InterPro; IPR013786; AcylCoA_DH/ox_N.
DR InterPro; IPR037069; AcylCoA_DH/ox_N_sf.
DR InterPro; IPR009100; AcylCoA_DH/oxidase_NM_dom.
DR Pfam; PF00441; Acyl-CoA_dh_1; 1.
DR Pfam; PF02771; Acyl-CoA_dh_N; 1.
DR SUPFAM; SSF47203; SSF47203; 1.
DR SUPFAM; SSF56645; SSF56645; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; FAD; Flavoprotein;
KW Nucleotide-binding; Oxidoreductase.
FT CHAIN 1..439
FT /note="Nitroalkane oxidase"
FT /id="PRO_0000418397"
FT ACT_SITE 402
FT /note="Proton acceptor"
FT /evidence="ECO:0000305|PubMed:11867731,
FT ECO:0000305|PubMed:12862464, ECO:0000305|PubMed:16430210"
FT BINDING 131..134
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT BINDING 139..141
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT BINDING 169..171
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT BINDING 304
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT BINDING 313..314
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT BINDING 375..379
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT BINDING 400..404
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000269|PubMed:16430210,
FT ECO:0000269|PubMed:17994768, ECO:0000269|PubMed:19926855"
FT MUTAGEN 276
FT /note="S->A: Decreases catalytic activity about tenfold."
FT /evidence="ECO:0000269|PubMed:19265437"
FT MUTAGEN 402
FT /note="D->E: Decreases enzyme activity about twentyfold."
FT /evidence="ECO:0000269|PubMed:11867731,
FT ECO:0000269|PubMed:12862464, ECO:0000269|PubMed:19265437"
FT MUTAGEN 402
FT /note="D->N: Almost abolishes enzyme activity towards
FT neutral nitroethane, but retains activity towards anionic
FT nitroethane."
FT /evidence="ECO:0000269|PubMed:11867731,
FT ECO:0000269|PubMed:12862464, ECO:0000269|PubMed:19265437"
FT MUTAGEN 409
FT /note="R->K: Reduces catalytic activity."
FT /evidence="ECO:0000269|PubMed:17994768"
FT HELIX 8..23
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 25..27
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 28..32
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 38..43
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 46..54
FT /evidence="ECO:0007829|PDB:2C12"
FT TURN 55..58
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 59..61
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 73..84
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 91..106
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 110..116
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 118..121
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 129..132
FT /evidence="ECO:0007829|PDB:2C12"
FT TURN 140..143
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 153..157
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 160..168
FT /evidence="ECO:0007829|PDB:2C12"
FT TURN 171..177
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 182..190
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 204..207
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 208..213
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 215..219
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 223..225
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 226..230
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 234..236
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 244..253
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 254..256
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 263..301
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 306..308
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 310..312
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 314..341
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 344..346
FT /evidence="ECO:0007829|PDB:2ZAF"
FT HELIX 348..377
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 379..382
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 388..395
FT /evidence="ECO:0007829|PDB:2C12"
FT TURN 396..400
FT /evidence="ECO:0007829|PDB:2C12"
FT STRAND 401..403
FT /evidence="ECO:0007829|PDB:2C12"
FT TURN 405..408
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 409..417
FT /evidence="ECO:0007829|PDB:2C12"
FT HELIX 426..429
FT /evidence="ECO:0007829|PDB:2C12"
SQ SEQUENCE 439 AA; 48162 MW; 9F156889F2A5901A CRC64;
MVDFKLSPSQ LEARRHAQAF ANTVLTKASA EYSTQKDQLS RFQATRPFYR EAVRHGLIKA
QVPIPLGGTM ESLVHESIIL EELFAVEPAT SITIVATALG LMPVILCDSP SLQEKFLKPF
ISGEGEPLAS LMHSEPNGTA NWLQKGGPGL QTTARKVGNE WVISGEKLWP SNSGGWDYKG
ADLACVVCRV SDDPSKPQDP NVDPATQIAV LLVTRETIAN NKKDAYQILG EPELAGHITT
SGPHTRFTEF HVPHENLLCT PGLKAQGLVE TAFAMSAALV GAMAIGTARA AFEEALVFAK
SDTRGGSKHI IEHQSVADKL IDCKIRLETS RLLVWKAVTT LEDEALEWKV KLEMAMQTKI
YTTDVAVECV IDAMKAVGMK SYAKDMSFPR LLNEVMCYPL FDGGNIGLRR RQMQRVMALE
DYEPWAATYG SSKVDKSRL
