NFSB_ECOLI
ID NFSB_ECOLI Reviewed; 217 AA.
AC P38489; P19575;
DT 01-OCT-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1994, sequence version 1.
DT 03-AUG-2022, entry version 174.
DE RecName: Full=Oxygen-insensitive NAD(P)H nitroreductase;
DE EC=1.-.-.-;
DE AltName: Full=Dihydropteridine reductase;
DE EC=1.5.1.34;
DE AltName: Full=FMN-dependent nitroreductase;
GN Name=nfsB {ECO:0000303|PubMed:8947835};
GN Synonyms=dprA, nfnB {ECO:0000303|PubMed:7813889}, nfsI, ntr;
GN OrderedLocusNames=b0578, JW0567;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12 / C600 / CR34 / ATCC 23724 / DSM 3925 / LMG 3041 / NCIB 10222;
RX PubMed=8947835; DOI=10.1093/oxfordjournals.jbchem.a021473;
RA Zenno S., Koike H., Tanokura M., Saigo K.;
RT "Gene cloning, purification, and characterization of NfsB, a minor oxygen-
RT insensitive nitroreductase from Escherichia coli, similar in biochemical
RT properties to FRase I, the major flavin reductase in Vibrio fischeri.";
RL J. Biochem. 120:736-744(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=B;
RX PubMed=7813889; DOI=10.1111/j.1574-6968.1994.tb07284.x;
RA Michael N.P., Brehm J.K., Anlezark G.M., Minton N.P.;
RT "Physical characterisation of the Escherichia coli B gene encoding
RT nitroreductase and its over-expression in Escherichia coli K12.";
RL FEMS Microbiol. Lett. 124:195-202(1994).
RN [3]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=8905232; DOI=10.1093/dnares/3.3.137;
RA Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K.,
RA Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S.,
RA Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H.,
RA Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G.,
RA Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M.,
RA Horiuchi T.;
RT "A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to
RT the 12.7-28.0 min region on the linkage map.";
RL DNA Res. 3:137-155(1996).
RN [4]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RA Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT "Sequence of minutes 4-25 of Escherichia coli.";
RL Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN [5]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA Shao Y.;
RT "The complete genome sequence of Escherichia coli K-12.";
RL Science 277:1453-1462(1997).
RN [6]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=16738553; DOI=10.1038/msb4100049;
RA Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT and W3110.";
RL Mol. Syst. Biol. 2:E1-E5(2006).
RN [7]
RP PROTEIN SEQUENCE OF 1-31 AND 139-180, AND CHARACTERIZATION.
RC STRAIN=B;
RX PubMed=1472094; DOI=10.1016/0006-2952(92)90671-5;
RA Anlezark G.M., Melton R.G., Sherwood R.F., Coles B., Friedlos F.,
RA Knox R.J.;
RT "The bioactivation of 5-(aziridin-1-yl)-2,4-dinitrobenzamide (CB1954) -- I.
RT Purification and properties of a nitroreductase enzyme from Escherichia
RT coli -- a potential enzyme for antibody-directed enzyme prodrug therapy
RT (ADEPT).";
RL Biochem. Pharmacol. 44:2289-2295(1992).
RN [8]
RP PROTEIN SEQUENCE OF 1-12.
RC STRAIN=K12 / EMG2;
RX PubMed=9298646; DOI=10.1002/elps.1150180807;
RA Link A.J., Robison K., Church G.M.;
RT "Comparing the predicted and observed properties of proteins encoded in the
RT genome of Escherichia coli K-12.";
RL Electrophoresis 18:1259-1313(1997).
RN [9]
RP PROTEIN SEQUENCE OF 1-20, AND CHARACTERIZATION AS A DIHYDROPTERIDINE
RP REDUCT.
RX PubMed=3060113;
RA Vasudevan S.G., Shaw D.C., Armarego W.L.F.;
RT "Dihydropteridine reductase from Escherichia coli.";
RL Biochem. J. 255:581-588(1988).
RN [10]
RP X-RAY CRYSTALLOGRAPHY (2.06 ANGSTROMS) IN COMPLEX WITH FMN, AND SUBUNIT.
RC STRAIN=B;
RX PubMed=11020276; DOI=10.1021/jm000159m;
RA Parkinson G.N., Skelly J.V., Neidle S.;
RT "Crystal structure of FMN-dependent nitroreductase from Escherichia coli B:
RT a prodrug-activating enzyme.";
RL J. Med. Chem. 43:3624-3631(2000).
RN [11]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH NICOTININC ACID AND
RP FMN, AND SUBUNIT.
RC STRAIN=K12 / DH5-alpha;
RX PubMed=11491290; DOI=10.1006/jmbi.2001.4653;
RA Lovering A.L., Hyde E.I., Searle P.F., White S.A.;
RT "The structure of Escherichia coli nitroreductase complexed with nicotinic
RT acid: three crystal forms at 1.7 A, 1.8 A and 2.4 A resolution.";
RL J. Mol. Biol. 309:203-213(2001).
RN [12]
RP X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEXES WITH FMN; DICOUMAROL AND
RP DINITROBENZAMIDE PRODRUGS, AND SUBUNIT.
RX PubMed=12954054; DOI=10.1021/jm030843b;
RA Johansson E., Parkinson G.N., Denny W.A., Neidle S.;
RT "Studies on the nitroreductase prodrug-activating system. Crystal
RT structures of complexes with the inhibitor dicoumarol and dinitrobenzamide
RT prodrugs and of the enzyme active form.";
RL J. Med. Chem. 46:4009-4020(2003).
RN [13]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH FMN; ACETATE AND THE
RP ANTIBIOTIC NITROFURAZONE, FUNCTION, ACTIVITY REGULATION, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=15684426; DOI=10.1074/jbc.m409652200;
RA Race P.R., Lovering A.L., Green R.M., Ossor A., White S.A., Searle P.F.,
RA Wrighton C.J., Hyde E.I.;
RT "Structural and mechanistic studies of Escherichia coli nitroreductase with
RT the antibiotic nitrofurazone. Reversed binding orientations in different
RT redox states of the enzyme.";
RL J. Biol. Chem. 280:13256-13264(2005).
CC -!- FUNCTION: Reduction of a variety of nitroaromatic compounds using NADH
CC (and to lesser extent NADPH) as source of reducing equivalents; two
CC electrons are transferred. Capable of reducing nitrofurazone, quinones
CC and the anti-tumor agent CB1954 (5-(aziridin-1-yl)-2,4-
CC dinitrobenzamide). The reduction of CB1954 results in the generation of
CC cytotoxic species. {ECO:0000269|PubMed:15684426}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5,6,7,8-tetrahydropteridine + NAD(+) = 6,7-dihydropteridine +
CC H(+) + NADH; Xref=Rhea:RHEA:17869, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:28889, ChEBI:CHEBI:30156, ChEBI:CHEBI:57540,
CC ChEBI:CHEBI:57945; EC=1.5.1.34;
CC -!- CATALYTIC ACTIVITY:
CC Reaction=5,6,7,8-tetrahydropteridine + NADP(+) = 6,7-dihydropteridine +
CC H(+) + NADPH; Xref=Rhea:RHEA:17865, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:28889, ChEBI:CHEBI:30156, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.5.1.34;
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC -!- ACTIVITY REGULATION: Subject to competitive inhibition by dicoumarol,
CC nicotinic acid and acetate with respect to NADH. Subject to
CC uncompetitive inhibition by dicoumarol, nicotinic acid and acetate with
CC respect to nitrofurazone and nitrofurantoin.
CC {ECO:0000269|PubMed:15684426}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=350 uM for NADH {ECO:0000269|PubMed:15684426};
CC KM=1850 uM for nitrofurazone {ECO:0000269|PubMed:15684426};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11020276,
CC ECO:0000269|PubMed:11491290, ECO:0000269|PubMed:12954054,
CC ECO:0000269|PubMed:15684426}.
CC -!- INTERACTION:
CC P38489; P38489: nfsB; NbExp=2; IntAct=EBI-909296, EBI-909296;
CC -!- SIMILARITY: Belongs to the nitroreductase family. {ECO:0000305}.
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DR EMBL; D25414; BAA05004.1; -; Genomic_DNA.
DR EMBL; U07860; AAC43263.1; -; Genomic_DNA.
DR EMBL; U82598; AAB40776.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73679.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA35218.1; -; Genomic_DNA.
DR PIR; I67685; I67685.
DR PIR; S01818; S01818.
DR RefSeq; NP_415110.1; NC_000913.3.
DR RefSeq; WP_000351487.1; NZ_SSZK01000024.1.
DR PDB; 1DS7; X-ray; 2.06 A; A/B=1-217.
DR PDB; 1ICR; X-ray; 1.70 A; A/B=1-217.
DR PDB; 1ICU; X-ray; 1.80 A; A/B/C/D=1-217.
DR PDB; 1ICV; X-ray; 2.40 A; A/B/C/D=1-217.
DR PDB; 1IDT; X-ray; 2.00 A; A/B=1-217.
DR PDB; 1OO5; X-ray; 2.50 A; A/B=1-217.
DR PDB; 1OO6; X-ray; 2.00 A; A/B=1-217.
DR PDB; 1OON; X-ray; 2.49 A; A/B=1-217.
DR PDB; 1OOQ; X-ray; 2.00 A; A/B=1-217.
DR PDB; 1YKI; X-ray; 1.70 A; A/B/C/D=1-217.
DR PDB; 1YLR; X-ray; 1.70 A; A/B=1-217.
DR PDB; 1YLU; X-ray; 2.00 A; A/B=1-217.
DR PDB; 3X21; X-ray; 3.00 A; A/B/C/D/E/F/G/H/I/J=1-217.
DR PDB; 3X22; X-ray; 2.00 A; A/B=1-217.
DR PDBsum; 1DS7; -.
DR PDBsum; 1ICR; -.
DR PDBsum; 1ICU; -.
DR PDBsum; 1ICV; -.
DR PDBsum; 1IDT; -.
DR PDBsum; 1OO5; -.
DR PDBsum; 1OO6; -.
DR PDBsum; 1OON; -.
DR PDBsum; 1OOQ; -.
DR PDBsum; 1YKI; -.
DR PDBsum; 1YLR; -.
DR PDBsum; 1YLU; -.
DR PDBsum; 3X21; -.
DR PDBsum; 3X22; -.
DR AlphaFoldDB; P38489; -.
DR SMR; P38489; -.
DR BioGRID; 4262913; 20.
DR BioGRID; 850145; 1.
DR DIP; DIP-10330N; -.
DR IntAct; P38489; 12.
DR STRING; 511145.b0578; -.
DR BindingDB; P38489; -.
DR ChEMBL; CHEMBL1075080; -.
DR DrugBank; DB04138; 5-[Bis(2-bromoethyl)amino]-N-(2,3-dioxopropyl)-2,4-dinitrobenzamide.
DR DrugBank; DB03228; 5-[Bis-2(Chloro-Ethyl)-Amino]-2,4-Dintro-Benzamide.
DR DrugBank; DB04392; Bishydroxy[2h-1-Benzopyran-2-One,1,2-Benzopyrone].
DR DrugBank; DB03247; Flavin mononucleotide.
DR DrugBank; DB04253; Tretazicar.
DR SWISS-2DPAGE; P38489; -.
DR jPOST; P38489; -.
DR PaxDb; P38489; -.
DR PRIDE; P38489; -.
DR EnsemblBacteria; AAC73679; AAC73679; b0578.
DR EnsemblBacteria; BAA35218; BAA35218; BAA35218.
DR GeneID; 945778; -.
DR KEGG; ecj:JW0567; -.
DR KEGG; eco:b0578; -.
DR PATRIC; fig|1411691.4.peg.1696; -.
DR EchoBASE; EB4146; -.
DR eggNOG; COG0778; Bacteria.
DR HOGENOM; CLU_070764_4_1_6; -.
DR InParanoid; P38489; -.
DR OMA; PWHFVVA; -.
DR PhylomeDB; P38489; -.
DR BioCyc; EcoCyc:DIHYDROPTERIREDUCT-MON; -.
DR BioCyc; MetaCyc:DIHYDROPTERIREDUCT-MON; -.
DR BRENDA; 1.7.1.B2; 2026.
DR EvolutionaryTrace; P38489; -.
DR PRO; PR:P38489; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0016020; C:membrane; HDA:UniProtKB.
DR GO; GO:0004155; F:6,7-dihydropteridine reductase activity; IDA:EcoCyc.
DR GO; GO:0010181; F:FMN binding; IDA:EcoCyc.
DR GO; GO:0042802; F:identical protein binding; IPI:IntAct.
DR GO; GO:0003955; F:NAD(P)H dehydrogenase (quinone) activity; IDA:EcoCyc.
DR GO; GO:0018545; F:NAD(P)H nitroreductase activity; IDA:EcoCyc.
DR GO; GO:0016491; F:oxidoreductase activity; IBA:GO_Central.
DR GO; GO:0046256; P:2,4,6-trinitrotoluene catabolic process; IMP:EcoCyc.
DR CDD; cd02149; NfsB-like; 1.
DR Gene3D; 3.40.109.10; -; 1.
DR InterPro; IPR033878; NfsB-like.
DR InterPro; IPR029479; Nitroreductase.
DR InterPro; IPR000415; Nitroreductase-like.
DR Pfam; PF00881; Nitroreductase; 1.
DR SUPFAM; SSF55469; SSF55469; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Direct protein sequencing; Flavoprotein; FMN; NAD; NADP;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..217
FT /note="Oxygen-insensitive NAD(P)H nitroreductase"
FT /id="PRO_0000072711"
FT BINDING 10..14
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11020276,
FT ECO:0000269|PubMed:11491290, ECO:0000269|PubMed:15684426"
FT BINDING 14
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:Q01234"
FT BINDING 41
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:Q01234"
FT BINDING 71
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11020276,
FT ECO:0000269|PubMed:11491290, ECO:0000269|PubMed:15684426"
FT BINDING 71
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:Q01234"
FT BINDING 74
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:Q01234"
FT BINDING 107
FT /ligand="NAD(+)"
FT /ligand_id="ChEBI:CHEBI:57540"
FT /evidence="ECO:0000250|UniProtKB:Q01234"
FT BINDING 165..166
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11020276,
FT ECO:0000269|PubMed:11491290, ECO:0000269|PubMed:15684426"
FT BINDING 205..207
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11020276,
FT ECO:0000269|PubMed:11491290, ECO:0000269|PubMed:15684426"
FT CONFLICT 5
FT /note="S -> C (in Ref. 9; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 10..12
FT /note="RHS -> CIV (in Ref. 9; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 19
FT /note="S -> M (in Ref. 9; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 21
FT /note="Missing (in Ref. 7; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 28
FT /note="E -> D (in Ref. 7; AA sequence)"
FT /evidence="ECO:0000305"
FT CONFLICT 180
FT /note="E -> I (in Ref. 7; AA sequence)"
FT /evidence="ECO:0000305"
FT HELIX 3..9
FT /evidence="ECO:0007829|PDB:1ICR"
FT STRAND 14..16
FT /evidence="ECO:0007829|PDB:1OO6"
FT HELIX 24..36
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 40..42
FT /evidence="ECO:0007829|PDB:1ICR"
FT STRAND 46..51
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 54..61
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 62..64
FT /evidence="ECO:0007829|PDB:3X22"
FT HELIX 66..71
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 72..77
FT /evidence="ECO:0007829|PDB:1ICR"
FT STRAND 78..89
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 92..104
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 111..130
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 135..156
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 169..176
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 178..181
FT /evidence="ECO:0007829|PDB:1ICR"
FT STRAND 183..192
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 199..201
FT /evidence="ECO:0007829|PDB:1ICR"
FT HELIX 210..213
FT /evidence="ECO:0007829|PDB:1ICR"
FT STRAND 214..217
FT /evidence="ECO:0007829|PDB:1ICR"
SQ SEQUENCE 217 AA; 23905 MW; A516CEFC3D46AEAC CRC64;
MDIISVALKR HSTKAFDASK KLTPEQAEQI KTLLQYSPSS TNSQPWHFIV ASTEEGKARV
AKSAAGNYVF NERKMLDASH VVVFCAKTAM DDVWLKLVVD QEDADGRFAT PEAKAANDKG
RKFFADMHRK DLHDDAEWMA KQVYLNVGNF LLGVAALGLD AVPIEGFDAA ILDAEFGLKE
KGYTSLVVVP VGHHSVEDFN ATLPKSRLPQ NITLTEV
