NFSA_ECOLI
ID NFSA_ECOLI Reviewed; 240 AA.
AC P17117;
DT 01-AUG-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1995, sequence version 2.
DT 03-AUG-2022, entry version 164.
DE RecName: Full=Oxygen-insensitive NADPH nitroreductase {ECO:0000305};
DE EC=1.-.-.- {ECO:0000269|PubMed:8755878};
DE AltName: Full=Modulator of drug activity A;
GN Name=nfsA; Synonyms=mda18, mdaA {ECO:0000303|PubMed:7568050}, ybjB;
GN OrderedLocusNames=b0851, JW0835;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=7568050; DOI=10.1073/pnas.92.19.8950;
RA Chatterjee P.K., Sternberg N.L.;
RT "A general genetic approach in Escherichia coli for determining the
RT mechanism(s) of action of tumoricidal agents: application to DMP 840, a
RT tumoricidal agent.";
RL Proc. Natl. Acad. Sci. U.S.A. 92:8950-8954(1995).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR,
RP AND BIOPHYSICOCHEMICAL PROPERTIES.
RC STRAIN=K12 / AB1157;
RX PubMed=8755878; DOI=10.1128/jb.178.15.4508-4514.1996;
RA Zenno S., Koike H., Kumar A.N., Jayaraman R., Tanokura M., Saigo K.;
RT "Biochemical characterization of NfsA, the Escherichia coli major
RT nitroreductase exhibiting a high amino acid sequence homology to Frp, a
RT Vibrio harveyi flavin oxidoreductase.";
RL J. Bacteriol. 178:4508-4514(1996).
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;
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 [5]
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 [6]
RP PRELIMINARY NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 134-240.
RC STRAIN=K12;
RX PubMed=2570347; DOI=10.1007/bf02464894;
RA Kang W.-K., Icho T., Isono S., Kitakawa M., Isono K.;
RT "Characterization of the gene rimK responsible for the addition of glutamic
RT acid residues to the C-terminus of ribosomal protein S6 in Escherichia coli
RT K12.";
RL Mol. Gen. Genet. 217:281-288(1989).
RN [7]
RP MUTAGENESIS.
RX PubMed=9440535; DOI=10.1128/jb.180.2.422-425.1998;
RA Zenno S., Kobori T., Tanokura M., Saigo K.;
RT "Conversion of NfsA, the major Escherichia coli nitroreductase, to a flavin
RT reductase with an activity similar to that of Frp, a flavin reductase in
RT Vibrio harveyi, by a single amino acid substitution.";
RL J. Bacteriol. 180:422-425(1998).
RN [8]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) IN COMPLEX WITH FMN, COFACTOR,
RP SUBUNIT, AND MUTAGENESIS OF ARG-203 AND ARG-208.
RX PubMed=11034992; DOI=10.1074/jbc.m002617200;
RA Kobori T., Sasaki H., Lee W.C., Zenno S., Saigo K., Murphy M.E.,
RA Tanokura M.;
RT "Structure and site-directed mutagenesis of a flavoprotein from Escherichia
RT coli that reduces nitrocompounds: alteration of pyridine nucleotide binding
RT by a single amino acid substitution.";
RL J. Biol. Chem. 276:2816-2823(2001).
CC -!- FUNCTION: Catalyzes the reduction of nitroaromatic compounds using
CC NADPH. Has a broad electron acceptor specificity. Reduces nitrofurazone
CC by a ping-pong bi-bi mechanism possibly to generate a two-electron
CC transfer product. Major oxygen-insensitive nitroreductase in E.coli.
CC {ECO:0000269|PubMed:8755878}.
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000269|PubMed:11034992, ECO:0000269|PubMed:8755878};
CC Note=Binds 1 FMN per monomer. {ECO:0000269|PubMed:11034992};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=11 uM for NADPH {ECO:0000269|PubMed:8755878};
CC KM=5.5 uM for nitrofurazone {ECO:0000269|PubMed:8755878};
CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:11034992}.
CC -!- INTERACTION:
CC P17117; P28630: holA; NbExp=3; IntAct=EBI-1120624, EBI-549153;
CC -!- SIMILARITY: Belongs to the flavin oxidoreductase frp family.
CC {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=CAA33867.1; Type=Frameshift; Evidence={ECO:0000305};
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DR EMBL; U18655; AAC43450.1; -; Genomic_DNA.
DR EMBL; D38308; BAA07425.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73938.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA35562.1; -; Genomic_DNA.
DR EMBL; X15859; CAA33867.1; ALT_FRAME; Genomic_DNA.
DR PIR; I80318; I80318.
DR RefSeq; NP_415372.1; NC_000913.3.
DR RefSeq; WP_000189159.1; NZ_LN832404.1.
DR PDB; 1F5V; X-ray; 1.70 A; A/B=1-240.
DR PDB; 7NB9; X-ray; 1.09 A; A=1-240.
DR PDB; 7NIY; X-ray; 1.03 A; A=1-240.
DR PDB; 7NMP; X-ray; 1.25 A; A=1-240.
DR PDB; 7NNX; X-ray; 1.70 A; A=1-240.
DR PDBsum; 1F5V; -.
DR PDBsum; 7NB9; -.
DR PDBsum; 7NIY; -.
DR PDBsum; 7NMP; -.
DR PDBsum; 7NNX; -.
DR AlphaFoldDB; P17117; -.
DR SMR; P17117; -.
DR BioGRID; 4259991; 28.
DR BioGRID; 849857; 4.
DR DIP; DIP-10334N; -.
DR IntAct; P17117; 5.
DR STRING; 511145.b0851; -.
DR DrugBank; DB03247; Flavin mononucleotide.
DR DrugBank; DB00698; Nitrofurantoin.
DR jPOST; P17117; -.
DR PaxDb; P17117; -.
DR PRIDE; P17117; -.
DR EnsemblBacteria; AAC73938; AAC73938; b0851.
DR EnsemblBacteria; BAA35562; BAA35562; BAA35562.
DR GeneID; 945483; -.
DR KEGG; ecj:JW0835; -.
DR KEGG; eco:b0851; -.
DR PATRIC; fig|1411691.4.peg.1427; -.
DR EchoBASE; EB1241; -.
DR eggNOG; COG0778; Bacteria.
DR HOGENOM; CLU_070764_0_2_6; -.
DR InParanoid; P17117; -.
DR OMA; GMCLGYP; -.
DR PhylomeDB; P17117; -.
DR BioCyc; EcoCyc:EG11261-MON; -.
DR BioCyc; MetaCyc:EG11261-MON; -.
DR BRENDA; 1.7.1.B3; 2026.
DR EvolutionaryTrace; P17117; -.
DR PRO; PR:P17117; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005829; C:cytosol; IDA:EcoCyc.
DR GO; GO:0034567; F:chromate reductase activity; IDA:EcoCyc.
DR GO; GO:0010181; F:FMN binding; IDA:EcoCyc.
DR GO; GO:0003955; F:NAD(P)H dehydrogenase (quinone) activity; IDA:EcoCyc.
DR GO; GO:0016657; F:oxidoreductase activity, acting on NAD(P)H, nitrogenous group as acceptor; IDA:EcoCyc.
DR CDD; cd02146; NfsA-like; 1.
DR Gene3D; 3.40.109.10; -; 1.
DR InterPro; IPR016446; Flavin_OxRdtase_Frp.
DR InterPro; IPR029479; Nitroreductase.
DR InterPro; IPR000415; Nitroreductase-like.
DR PANTHER; PTHR43425; PTHR43425; 1.
DR Pfam; PF00881; Nitroreductase; 1.
DR PIRSF; PIRSF005426; Frp; 1.
DR SUPFAM; SSF55469; SSF55469; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Flavoprotein; FMN; NADP; Oxidoreductase; Reference proteome.
FT CHAIN 1..240
FT /note="Oxygen-insensitive NADPH nitroreductase"
FT /id="PRO_0000205510"
FT BINDING 11..15
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11034992,
FT ECO:0007744|PDB:1F5V"
FT BINDING 39
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11034992,
FT ECO:0007744|PDB:1F5V"
FT BINDING 67
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11034992,
FT ECO:0007744|PDB:1F5V"
FT BINDING 128..131
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11034992,
FT ECO:0007744|PDB:1F5V"
FT BINDING 167..169
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000269|PubMed:11034992,
FT ECO:0007744|PDB:1F5V"
FT MUTAGEN 203
FT /note="R->A: Strong decrease in activity."
FT /evidence="ECO:0000269|PubMed:11034992"
FT MUTAGEN 208
FT /note="R->A: No change in activity."
FT /evidence="ECO:0000269|PubMed:11034992"
FT HELIX 3..9
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 24..35
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 40..42
FT /evidence="ECO:0007829|PDB:7NIY"
FT STRAND 46..50
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 54..63
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 69..72
FT /evidence="ECO:0007829|PDB:7NIY"
FT STRAND 73..82
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 84..89
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 98..121
FT /evidence="ECO:0007829|PDB:7NIY"
FT STRAND 125..129
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 130..135
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 136..142
FT /evidence="ECO:0007829|PDB:7NIY"
FT STRAND 149..158
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 172..175
FT /evidence="ECO:0007829|PDB:7NIY"
FT STRAND 178..180
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 186..202
FT /evidence="ECO:0007829|PDB:7NIY"
FT STRAND 203..206
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 212..219
FT /evidence="ECO:0007829|PDB:7NIY"
FT HELIX 228..234
FT /evidence="ECO:0007829|PDB:7NIY"
SQ SEQUENCE 240 AA; 26801 MW; 3B389FCDB86DED1D CRC64;
MTPTIELICG HRSIRHFTDE PISEAQREAI INSARATSSS SFLQCSSIIR ITDKALREEL
VTLTGGQKHV AQAAEFWVFC ADFNRHLQIC PDAQLGLAEQ LLLGVVDTAM MAQNALIAAE
SLGLGGVYIG GLRNNIEAVT KLLKLPQHVL PLFGLCLGWP ADNPDLKPRL PASILVHENS
YQPLDKGALA QYDEQLAEYY LTRGSNNRRD TWSDHIRRTI IKESRPFILD YLHKQGWATR
