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Protein

Oxygen-insensitive NAD(P)H nitroreductase

Gene

nfsB

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Reduction of a variety of nitroaromatic compounds using NADH (and to lesser extent NADPH) as source of reducing equivalents; two electrons are transferred. Capable of reducing nitrofurazone, quinones and the anti-tumor agent CB1954 (5-(aziridin-1-yl)-2,4-dinitrobenzamide). The reduction of CB1954 results in the generation of cytotoxic species.1 Publication

Catalytic activityi

A 5,6,7,8-tetrahydropteridine + NAD(P)+ = a 6,7-dihydropteridine + NAD(P)H.

Cofactori

Enzyme regulationi

Subject to competitive inhibition by dicoumarol, nicotinic acid and acetate with respect to NADH. Subject to uncompetitive inhibition by dicoumarol, nicotinic acid and acetate with respect to nitrofurazone and nitrofurantoin.1 Publication

Kineticsi

  1. KM=350 µM for NADH1 Publication
  2. KM=1850 µM for nitrofurazone1 Publication

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Binding sitei41 – 411NAD or NADP; via amide nitrogen
    Binding sitei71 – 711FMN3 Publications

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi10 – 145FMN3 Publications
    Nucleotide bindingi153 – 1586NAD or NADPBy similarity
    Nucleotide bindingi165 – 1662FMN3 Publications
    Nucleotide bindingi205 – 2073FMN3 Publications

    GO - Molecular functioni

    • 6,7-dihydropteridine reductase activity Source: EcoCyc
    • FAD binding Source: EcoCyc
    • FMN binding Source: EcoCyc
    • identical protein binding Source: IntAct
    • NAD(P)H nitroreductase activity Source: EcoCyc
    Complete GO annotation...

    Keywords - Molecular functioni

    Oxidoreductase

    Keywords - Ligandi

    Flavoprotein, FMN, NAD, NADP

    Enzyme and pathway databases

    BioCyciEcoCyc:DIHYDROPTERIREDUCT-MONOMER.
    ECOL316407:JW0567-MONOMER.
    MetaCyc:DIHYDROPTERIREDUCT-MONOMER.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    Oxygen-insensitive NAD(P)H nitroreductase (EC:1.-.-.-)
    Alternative name(s):
    Dihydropteridine reductase (EC:1.5.1.34)
    FMN-dependent nitroreductase
    Gene namesi
    Name:nfsB1 Publication
    Synonyms:dprA, nfnB1 Publication, nfsI, ntr
    Ordered Locus Names:b0578, JW0567
    OrganismiEscherichia coli (strain K12)
    Taxonomic identifieri83333 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
    ProteomesiUP000000318 Componenti: Chromosome UP000000625 Componenti: Chromosome

    Organism-specific databases

    EcoGeneiEG20151. nfsB.

    Subcellular locationi

    GO - Cellular componenti

    • cytosol Source: UniProtKB
    • membrane Source: UniProtKB
    Complete GO annotation...

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 217217Oxygen-insensitive NAD(P)H nitroreductasePRO_0000072711Add
    BLAST

    Proteomic databases

    PaxDbiP38489.
    PRIDEiP38489.

    2D gel databases

    SWISS-2DPAGEP38489.

    Interactioni

    Subunit structurei

    Homodimer.4 Publications

    Binary interactionsi

    WithEntry#Exp.IntActNotes
    itself2EBI-909296,EBI-909296

    Protein-protein interaction databases

    DIPiDIP-10330N.
    IntActiP38489. 12 interactions.
    STRINGi511145.b0578.

    Structurei

    Secondary structure

    1
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Helixi3 – 97Combined sources
    Beta strandi14 – 163Combined sources
    Helixi24 – 3613Combined sources
    Helixi40 – 423Combined sources
    Beta strandi46 – 516Combined sources
    Helixi54 – 618Combined sources
    Helixi62 – 643Combined sources
    Helixi66 – 716Combined sources
    Helixi72 – 776Combined sources
    Beta strandi78 – 8912Combined sources
    Helixi92 – 10413Combined sources
    Helixi111 – 13020Combined sources
    Helixi135 – 15622Combined sources
    Helixi169 – 1768Combined sources
    Helixi178 – 1814Combined sources
    Beta strandi183 – 19210Combined sources
    Helixi199 – 2013Combined sources
    Helixi210 – 2134Combined sources
    Beta strandi214 – 2174Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1DS7X-ray2.06A/B1-217[»]
    1ICRX-ray1.70A/B1-217[»]
    1ICUX-ray1.80A/B/C/D1-217[»]
    1ICVX-ray2.40A/B/C/D1-217[»]
    1IDTX-ray2.00A/B1-217[»]
    1OO5X-ray2.50A/B1-217[»]
    1OO6X-ray2.00A/B1-217[»]
    1OONX-ray2.49A/B1-217[»]
    1OOQX-ray2.00A/B1-217[»]
    1YKIX-ray1.70A/B/C/D1-217[»]
    1YLRX-ray1.70A/B1-217[»]
    1YLUX-ray2.00A/B1-217[»]
    3X21X-ray3.00A/B/C/D/E/F/G/H/I/J1-217[»]
    ProteinModelPortaliP38489.
    SMRiP38489. Positions 1-217.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP38489.

    Family & Domainsi

    Sequence similaritiesi

    Belongs to the nitroreductase family.Curated

    Phylogenomic databases

    eggNOGiCOG0778.
    HOGENOMiHOG000146740.
    InParanoidiP38489.
    KOiK10679.
    OMAiKRRYTTK.
    OrthoDBiEOG65QWJJ.
    PhylomeDBiP38489.

    Family and domain databases

    Gene3Di3.40.109.10. 1 hit.
    InterProiIPR029479. Nitroreductase.
    IPR000415. Nitroreductase-like.
    [Graphical view]
    PfamiPF00881. Nitroreductase. 1 hit.
    [Graphical view]
    SUPFAMiSSF55469. SSF55469. 1 hit.

    Sequencei

    Sequence statusi: Complete.

    P38489-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MDIISVALKR HSTKAFDASK KLTPEQAEQI KTLLQYSPSS TNSQPWHFIV
    60 70 80 90 100
    ASTEEGKARV AKSAAGNYVF NERKMLDASH VVVFCAKTAM DDVWLKLVVD
    110 120 130 140 150
    QEDADGRFAT PEAKAANDKG RKFFADMHRK DLHDDAEWMA KQVYLNVGNF
    160 170 180 190 200
    LLGVAALGLD AVPIEGFDAA ILDAEFGLKE KGYTSLVVVP VGHHSVEDFN
    210
    ATLPKSRLPQ NITLTEV
    Length:217
    Mass (Da):23,905
    Last modified:October 1, 1994 - v1
    Checksum:iA516CEFC3D46AEAC
    GO

    Experimental Info

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Sequence conflicti5 – 51S → C AA sequence (PubMed:3060113).Curated
    Sequence conflicti10 – 123RHS → CIV AA sequence (PubMed:3060113).Curated
    Sequence conflicti19 – 191S → M AA sequence (PubMed:3060113).Curated
    Sequence conflicti21 – 211Missing AA sequence (PubMed:1472094).Curated
    Sequence conflicti28 – 281E → D AA sequence (PubMed:1472094).Curated
    Sequence conflicti180 – 1801E → I AA sequence (PubMed:1472094).Curated

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D25414 Genomic DNA. Translation: BAA05004.1.
    U07860 Genomic DNA. Translation: AAC43263.1.
    U82598 Genomic DNA. Translation: AAB40776.1.
    U00096 Genomic DNA. Translation: AAC73679.1.
    AP009048 Genomic DNA. Translation: BAA35218.1.
    PIRiI67685.
    S01818.
    RefSeqiNP_415110.1. NC_000913.3.
    WP_000351487.1. NZ_CP010445.1.

    Genome annotation databases

    EnsemblBacteriaiAAC73679; AAC73679; b0578.
    BAA35218; BAA35218; BAA35218.
    GeneIDi945778.
    KEGGieco:b0578.
    PATRICi32116324. VBIEscCol129921_0602.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    D25414 Genomic DNA. Translation: BAA05004.1.
    U07860 Genomic DNA. Translation: AAC43263.1.
    U82598 Genomic DNA. Translation: AAB40776.1.
    U00096 Genomic DNA. Translation: AAC73679.1.
    AP009048 Genomic DNA. Translation: BAA35218.1.
    PIRiI67685.
    S01818.
    RefSeqiNP_415110.1. NC_000913.3.
    WP_000351487.1. NZ_CP010445.1.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1DS7X-ray2.06A/B1-217[»]
    1ICRX-ray1.70A/B1-217[»]
    1ICUX-ray1.80A/B/C/D1-217[»]
    1ICVX-ray2.40A/B/C/D1-217[»]
    1IDTX-ray2.00A/B1-217[»]
    1OO5X-ray2.50A/B1-217[»]
    1OO6X-ray2.00A/B1-217[»]
    1OONX-ray2.49A/B1-217[»]
    1OOQX-ray2.00A/B1-217[»]
    1YKIX-ray1.70A/B/C/D1-217[»]
    1YLRX-ray1.70A/B1-217[»]
    1YLUX-ray2.00A/B1-217[»]
    3X21X-ray3.00A/B/C/D/E/F/G/H/I/J1-217[»]
    ProteinModelPortaliP38489.
    SMRiP38489. Positions 1-217.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    DIPiDIP-10330N.
    IntActiP38489. 12 interactions.
    STRINGi511145.b0578.

    Chemistry

    ChEMBLiCHEMBL1075080.

    2D gel databases

    SWISS-2DPAGEP38489.

    Proteomic databases

    PaxDbiP38489.
    PRIDEiP38489.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiAAC73679; AAC73679; b0578.
    BAA35218; BAA35218; BAA35218.
    GeneIDi945778.
    KEGGieco:b0578.
    PATRICi32116324. VBIEscCol129921_0602.

    Organism-specific databases

    EchoBASEiEB4146.
    EcoGeneiEG20151. nfsB.

    Phylogenomic databases

    eggNOGiCOG0778.
    HOGENOMiHOG000146740.
    InParanoidiP38489.
    KOiK10679.
    OMAiKRRYTTK.
    OrthoDBiEOG65QWJJ.
    PhylomeDBiP38489.

    Enzyme and pathway databases

    BioCyciEcoCyc:DIHYDROPTERIREDUCT-MONOMER.
    ECOL316407:JW0567-MONOMER.
    MetaCyc:DIHYDROPTERIREDUCT-MONOMER.

    Miscellaneous databases

    EvolutionaryTraceiP38489.
    PROiP38489.

    Family and domain databases

    Gene3Di3.40.109.10. 1 hit.
    InterProiIPR029479. Nitroreductase.
    IPR000415. Nitroreductase-like.
    [Graphical view]
    PfamiPF00881. Nitroreductase. 1 hit.
    [Graphical view]
    SUPFAMiSSF55469. SSF55469. 1 hit.
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Gene cloning, purification, and characterization of NfsB, a minor oxygen-insensitive nitroreductase from Escherichia coli, similar in biochemical properties to FRase I, the major flavin reductase in Vibrio fischeri."
      Zenno S., Koike H., Tanokura M., Saigo K.
      J. Biochem. 120:736-744(1996) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: K12 / C600 / ATCC 23724 / DSM 3925 / LMG 3041 / NCIB 10222.
    2. "Physical characterisation of the Escherichia coli B gene encoding nitroreductase and its over-expression in Escherichia coli K12."
      Michael N.P., Brehm J.K., Anlezark G.M., Minton N.P.
      FEMS Microbiol. Lett. 124:195-202(1994) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: B.
    3. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
    4. "Sequence of minutes 4-25 of Escherichia coli."
      Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M., Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D., Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.
      Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    5. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    6. "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."
      Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.
      Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
    7. "The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) -- I. Purification and properties of a nitroreductase enzyme from Escherichia coli -- a potential enzyme for antibody-directed enzyme prodrug therapy (ADEPT)."
      Anlezark G.M., Melton R.G., Sherwood R.F., Coles B., Friedlos F., Knox R.J.
      Biochem. Pharmacol. 44:2289-2295(1992) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 1-31 AND 139-180, CHARACTERIZATION.
      Strain: B.
    8. "Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12."
      Link A.J., Robison K., Church G.M.
      Electrophoresis 18:1259-1313(1997) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 1-12.
      Strain: K12 / EMG2.
    9. "Dihydropteridine reductase from Escherichia coli."
      Vasudevan S.G., Shaw D.C., Armarego W.L.F.
      Biochem. J. 255:581-588(1988) [PubMed] [Europe PMC] [Abstract]
      Cited for: PROTEIN SEQUENCE OF 1-20, CHARACTERIZATION AS A DIHYDROPTERIDINE REDUCT.
    10. "Crystal structure of FMN-dependent nitroreductase from Escherichia coli B: a prodrug-activating enzyme."
      Parkinson G.N., Skelly J.V., Neidle S.
      J. Med. Chem. 43:3624-3631(2000) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.06 ANGSTROMS) IN COMPLEX WITH FMN, SUBUNIT.
      Strain: B.
    11. "The structure of Escherichia coli nitroreductase complexed with nicotinic acid: three crystal forms at 1.7 A, 1.8 A and 2.4 A resolution."
      Lovering A.L., Hyde E.I., Searle P.F., White S.A.
      J. Mol. Biol. 309:203-213(2001) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH NICOTININC ACID AND FMN, SUBUNIT.
      Strain: K12 / DH5-alpha.
    12. "Studies on the nitroreductase prodrug-activating system. Crystal structures of complexes with the inhibitor dicoumarol and dinitrobenzamide prodrugs and of the enzyme active form."
      Johansson E., Parkinson G.N., Denny W.A., Neidle S.
      J. Med. Chem. 46:4009-4020(2003) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEXES WITH FMN; DICOUMAROL AND DINITROBENZAMIDE PRODRUGS, SUBUNIT.
    13. "Structural and mechanistic studies of Escherichia coli nitroreductase with the antibiotic nitrofurazone. Reversed binding orientations in different redox states of the enzyme."
      Race P.R., Lovering A.L., Green R.M., Ossor A., White S.A., Searle P.F., Wrighton C.J., Hyde E.I.
      J. Biol. Chem. 280:13256-13264(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH FMN; ACETATE AND THE ANTIBIOTIC NITROFURAZONE, FUNCTION, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES.

    Entry informationi

    Entry nameiNFSB_ECOLI
    AccessioniPrimary (citable) accession number: P38489
    Secondary accession number(s): P19575
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: October 1, 1994
    Last sequence update: October 1, 1994
    Last modified: July 22, 2015
    This is version 132 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Complete proteome, Direct protein sequencing, Reference proteome

    Documents

    1. Escherichia coli
      Escherichia coli (strain K12): entries and cross-references to EcoGene
    2. PDB cross-references
      Index of Protein Data Bank (PDB) cross-references
    3. SIMILARITY comments
      Index of protein domains and families

    External Data

    Dasty 3

    Similar proteinsi

    Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
    100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into Uniref entry.
    90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
    50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.