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UniProtKB/Swiss-Prot entry P38489


[Entry info] [Name and origin] [References] [Comments] [Cross-references] [Keywords] [Features] [Sequence] [Tools]

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Entry information
Entry name NFNB_ECOLI
Primary accession number P38489
Secondary accession number P19575
Integrated into Swiss-Prot on October 1, 1994
Sequence was last modified on October 1, 1994 (Sequence version 1)
Annotations were last modified on    July 22, 2008 (Entry version 77)
Name and origin of the protein
Protein name Oxygen-insensitive NAD(P)H nitroreductase
Synonyms EC 1.-.-.-
FMN-dependent nitroreductase
Dihydropteridine reductase
EC 1.5.1.34
Gene name
Name: nfnB
Synonyms: dprA, nfsB, nfsI, ntr
OrderedLocusNames: b0578, JW0567
From
Escherichia coli (strain K12) [TaxID: 83333] [HAMAP proteome]
Taxonomy Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacteriales; Enterobacteriaceae; Escherichia.
Protein existence 1: Evidence at protein level;
References
[1]
NUCLEOTIDE SEQUENCE [GENOMIC DNA].
STRAIN=K12 / C600 / ATCC 23724 / DSM 3925 / LMG 3041 / NCIB 10222;
PubMed=8947835 [NCBI, ExPASy, EBI, Israel, Japan]
Zenno S., Koike H., Tanokura M., Saigo K.;
"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.";
J. Biochem. 120:736-744(1996).
[2]
NUCLEOTIDE SEQUENCE [GENOMIC DNA].
STRAIN=B;
DOI=10.1016/0378-1097(94)90249-6; PubMed=7813889 [NCBI, ExPASy, EBI, Israel, Japan]
Michael N.P., Brehm J.K., Anlezark G.M., Minton N.P.;
"Physical characterisation of the Escherichia coli B gene encoding nitroreductase and its over-expression in Escherichia coli K12.";
FEMS Microbiol. Lett. 124:195-202(1994).
[3]
NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
DOI=10.1093/dnares/3.3.137; PubMed=8905232 [NCBI, ExPASy, EBI, Israel, Japan]
Oshima T., Aiba H., Baba T., Fujita K., Hayashi K., Honjo A., Ikemoto K., Inada T., Itoh T., Kajihara M., Kanai K., Kashimoto K., Kimura S., Kitagawa M., Makino K., Masuda S., Miki T., Mizobuchi K., Mori H., Motomura K., Nakamura Y., Nashimoto H., Nishio Y., Saito N., Sampei G., Seki Y., Tagami H., Takemoto K., Wada C., Yamamoto Y., Yano M., Horiuchi T.;
"A 718-kb DNA sequence of the Escherichia coli K-12 genome corresponding to the 12.7-28.0 min region on the linkage map.";
DNA Res. 3:137-155(1996).
[4]
NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
STRAIN=K12 / MG1655 / ATCC 47076;
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.;
"Sequence of minutes 4-25 of Escherichia coli.";
Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
[5]
NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
STRAIN=K12 / MG1655 / ATCC 47076;
DOI=10.1126/science.277.5331.1453; PubMed=9278503 [NCBI, ExPASy, EBI, Israel, Japan]
Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.;
"The complete genome sequence of Escherichia coli K-12.";
Science 277:1453-1474(1997).
[6]
NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
DOI=10.1038/msb4100049; PubMed=16738553 [NCBI, ExPASy, EBI, Israel, Japan]
Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
"Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110.";
Mol. Syst. Biol. 2:E1-E5(2006).
[7]
PROTEIN SEQUENCE OF 1-31 AND 139-180, AND CHARACTERIZATION.
STRAIN=B;
DOI=10.1016/0006-2952(92)90671-5; PubMed=1472094 [NCBI, ExPASy, EBI, Israel, Japan]
Anlezark G.M., Melton R.G., Sherwood R.F., Coles B., Friedlos F., Knox R.J.;
"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).";
Biochem. Pharmacol. 44:2289-2295(1992).
[8]
PROTEIN SEQUENCE OF 1-12.
STRAIN=K12 / EMG2;
PubMed=9298646 [NCBI, ExPASy, EBI, Israel, Japan]
Link A.J., Robison K., Church G.M.;
"Comparing the predicted and observed properties of proteins encoded in the genome of Escherichia coli K-12.";
Electrophoresis 18:1259-1313(1997).
[9]
PROTEIN SEQUENCE OF 1-20, AND CHARACTERIZATION AS A DIHYDROPTERIDINE REDUCT.
PubMed=3060113 [NCBI, ExPASy, EBI, Israel, Japan]
Vasudevan S.G., Shaw D.C., Armarego W.L.F.;
"Dihydropteridine reductase from Escherichia coli.";
Biochem. J. 255:581-588(1988).
[10]
X-RAY CRYSTALLOGRAPHY (2.06 ANGSTROMS) IN COMPLEX WITH FMN, AND SUBUNIT.
STRAIN=B;
DOI=10.1021/jm000159m; PubMed=11020276 [NCBI, ExPASy, EBI, Israel, Japan]
Parkinson G.N., Skelly J.V., Neidle S.;
"Crystal structure of FMN-dependent nitroreductase from Escherichia coli B: a prodrug-activating enzyme.";
J. Med. Chem. 43:3624-3631(2000).
[11]
X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH NICOTININC ACID AND FMN, AND SUBUNIT.
STRAIN=K12 / DH5-alpha;
DOI=10.1006/jmbi.2001.4653; PubMed=11491290 [NCBI, ExPASy, EBI, Israel, Japan]
Lovering A.L., Hyde E.I., Searle P.F., White S.A.;
"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.";
J. Mol. Biol. 309:203-213(2001).
[12]
X-RAY CRYSTALLOGRAPHY (2.0 ANGSTROMS) IN COMPLEXES WITH FMN; DICOUMAROL AND DINITROBENZAMIDE PRODRUGS, AND SUBUNIT.
DOI=10.1021/jm030843b; PubMed=12954054 [NCBI, ExPASy, EBI, Israel, Japan]
Johansson E., Parkinson G.N., Denny W.A., Neidle S.;
"Studies on the nitroreductase prodrug-activating system. Crystal structures of complexes with the inhibitor dicoumarol and dinitrobenzamide prodrugs and of the enzyme active form.";
J. Med. Chem. 46:4009-4020(2003).
[13]
X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH FMN; ACETATE AND THE ANTIBIOTIC NITROFURAZONE, FUNCTION, ENZYME REGULATION, AND BIOPHYSICOCHEMICAL PROPERTIES.
DOI=10.1074/jbc.M409652200; PubMed=15684426 [NCBI, ExPASy, EBI, Israel, Japan]
Race P.R., Lovering A.L., Green R.M., Ossor A., White S.A., Searle P.F., Wrighton C.J., Hyde E.I.;
"Structural and mechanistic studies of Escherichia coli nitroreductase with the antibiotic nitrofurazone. Reversed binding orientations in different redox states of the enzyme.";
J. Biol. Chem. 280:13256-13264(2005).
Comments
  • FUNCTION: 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.
  • CATALYTIC ACTIVITY: A 5,6,7,8-tetrahydropteridine + NAD(P)+ = a 6,7-dihydropteridine + NAD(P)H.
  • COFACTOR: FMN.
  • ENZYME REGULATION: 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.
  • BIOPHYSICOCHEMICAL PROPERTIES:
    Kinetic parameters:   KM=350 µM for NADH;
    KM=1850 µM for nitrofurazone;
  • SUBUNIT: Homodimer.
  • SIMILARITY: Belongs to the nitroreductase family.
Copyright
Copyrighted by the UniProt Consortium, see http://www.uniprot.org/terms. Distributed under the Creative Commons Attribution-NoDerivs License.
Cross-references
Sequence databases
EMBL
D25414; BAA05004.1; -; Genomic_DNA.[EMBL / GenBank / DDBJ] [CoDingSequence]
U07860; AAC43263.1; -; Genomic_DNA.[EMBL / GenBank / DDBJ] [CoDingSequence]
U82598; AAB40776.1; -; Genomic_DNA.[EMBL / GenBank / DDBJ] [CoDingSequence]
U00096; AAC73679.1; -; Genomic_DNA.[EMBL / GenBank / DDBJ] [CoDingSequence]
AP009048; BAA35218.1; -; Genomic_DNA.[EMBL / GenBank / DDBJ] [CoDingSequence]
PIR I67685; I67685.
S01818; S01818.
RefSeq AP_001223.1; -.
NP_415110.1; -.
3D structure databases
PDB
1DS7; X-ray; 2.06 A; A/B=1-217.[ExPASy / RCSB / EBI]
1ICR; X-ray; 1.70 A; A/B=1-217.[ExPASy / RCSB / EBI]
1ICU; X-ray; 1.80 A; A/B/C/D=1-217.[ExPASy / RCSB / EBI]
1ICV; X-ray; 2.40 A; A/B/C/D=1-217.[ExPASy / RCSB / EBI]
1IDT; X-ray; 2.00 A; A/B=1-217.[ExPASy / RCSB / EBI]
1OO5; X-ray; 2.50 A; A/B=1-217.[ExPASy / RCSB / EBI]
1OO6; X-ray; 2.00 A; A/B=1-217.[ExPASy / RCSB / EBI]
1OON; X-ray; 2.49 A; A/B=1-217.[ExPASy / RCSB / EBI]
1OOQ; X-ray; 2.00 A; A/B=1-217.[ExPASy / RCSB / EBI]
1YKI; X-ray; 1.70 A; A/B/C/D=1-217.[ExPASy / RCSB / EBI]
1YLR; X-ray; 1.70 A; A/B=1-217.[ExPASy / RCSB / EBI]
1YLU; X-ray; 2.00 A; A/B=1-217.[ExPASy / RCSB / EBI]
Detailed list of linked structures.
PDBsum 1DS7; -.
1ICR; -.
1ICU; -.
1ICV; -.
1IDT; -.
1OO5; -.
1OO6; -.
1OON; -.
1OOQ; -.
1YKI; -.
1YLR; -.
1YLU; -.
ModBase P38489.
Protein-protein interaction databases
DIP DIP:10330N; -.
IntAct P38489; -.
Enzyme and pathway databases
BioCyc EcoCyc:DIHYDROPTERIREDUCT-MON; -.
2D gel databases
SWISS-2DPAGE P38489; -.
Organism-specific databases
EchoBASE EB4146; -.
EcoGene EG20151; nfnB.
Ontologies
GO
GO:0005624; Cellular component: membrane fraction (inferred from direct assay from UniProtKB).
QuickGo view.
Family and domain databases
InterPro IPR000415; Nitroreductase.
Graphical view of domain structure.
Pfam PF00881; Nitroreductase; 1.
Pfam graphical view of domain structure.
BLOCKS P38489.
Genome annotation databases
GeneID 945778; -.
GenomeReviews U00096_GR; b0578.
AP009048_GR; JW0567.
KEGG ecj:JW0567; -.
eco:b0578; -.
Phylogenomic databases
HOGENOM P38489; -.
Other
DrugBank DB01093; Dimethyl sulfoxide.
Genome annotation databases
CMR P38489; b0578.
Other
ProtoNet P38489.
UniRef View cluster of proteins with at least 50% / 90% / 100% identity.
Keywords
3D-structure; Complete proteome; Direct protein sequencing; Flavoprotein; FMN; NAD; NADP; Oxidoreductase.
Features
SEVIEWER logo Feature table viewer
KeyFrom   To Length Description FTId
CHAIN   1   217  217     Oxygen-insensitive NAD(P)H nitroreductase. PRO_0000072711
NP_BIND   10    14  5     FMN. 
NP_BIND   153   158  6     NAD or NADP (By similarity). 
NP_BIND   165   166  2     FMN. 
NP_BIND   205   207  3     FMN. 
BINDING   41    41        NAD or NADP; via amide nitrogen. 
BINDING   71    71        FMN. 
CONFLICT   5     5        S -> C (in Ref. 9; AA sequence). 
CONFLICT   10    12        RHS -> CIV (in Ref. 9; AA sequence). 
CONFLICT   19    19        S -> M (in Ref. 9; AA sequence). 
CONFLICT   21    21        Missing (in Ref. 7; AA sequence). 
CONFLICT   28    28        E -> D (in Ref. 7; AA sequence). 
CONFLICT   180   180        E -> I (in Ref. 7; AA sequence). 
HELIX   3     9  7      
HELIX   24    35  12      
HELIX   40    42  3      
STRAND   46    51  6      
HELIX   54    61  8      
HELIX   66    71  6      
HELIX   72    77  6      
STRAND   78    89  12      
HELIX   92   104  13      
HELIX   111   129  19      
TURN   130   132  3      
HELIX   135   157  23      
HELIX   169   175  7      
HELIX   179   181  3      
STRAND   183   192  10      
HELIX   199   201  3      
HELIX   210   213  4      
STRAND   214   216  3      
Sequence information
Length: 217 AA [This is the length of the unprocessed precursor] Molecular weight: 23905 Da [This is the MW of the unprocessed precursor] CRC64: A516CEFC3D46AEAC [This is a checksum on the sequence]
        10         20         30         40         50         60 
MDIISVALKR HSTKAFDASK KLTPEQAEQI KTLLQYSPSS TNSQPWHFIV ASTEEGKARV 

        70         80         90        100        110        120 
AKSAAGNYVF NERKMLDASH VVVFCAKTAM DDVWLKLVVD QEDADGRFAT PEAKAANDKG 

       130        140        150        160        170        180 
RKFFADMHRK DLHDDAEWMA KQVYLNVGNF LLGVAALGLD AVPIEGFDAA ILDAEFGLKE 

       190        200        210 
KGYTSLVVVP VGHHSVEDFN ATLPKSRLPQ NITLTEV 

P38489 in FASTA format

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View entry in raw text format (no links)
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