NEMA_ECOLI
ID NEMA_ECOLI Reviewed; 365 AA.
AC P77258; Q2MB63;
DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot.
DT 01-FEB-1997, sequence version 1.
DT 03-AUG-2022, entry version 144.
DE RecName: Full=N-ethylmaleimide reductase {ECO:0000303|PubMed:9013822};
DE Short=NEM reductase {ECO:0000303|PubMed:9013822};
DE EC=1.3.1.- {ECO:0000269|PubMed:23506073, ECO:0000269|PubMed:9013822};
DE AltName: Full=N-ethylmaleimide reducing enzyme;
GN Name=nemA {ECO:0000303|PubMed:9013822}; Synonyms=ydhN;
GN OrderedLocusNames=b1650, JW1642;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND CATALYTIC ACTIVITY.
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=9013822; DOI=10.1248/bpb.20.110;
RA Miura K., Tomioka Y., Suzuki H., Yonezawa M., Hishinuma T., Mizugaki M.;
RT "Molecular cloning of the nemA gene encoding N-ethylmaleimide reductase
RT from Escherichia coli.";
RL Biol. Pharm. Bull. 20:110-112(1997).
RN [2]
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 [3]
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 [4]
RP FUNCTION.
RC STRAIN=K12 / JM109 / ATCC 53323;
RX PubMed=15184158; DOI=10.1128/aem.70.6.3566-3574.2004;
RA Williams R.E., Rathbone D.A., Scrutton N.S., Bruce N.C.;
RT "Biotransformation of explosives by the old yellow enzyme family of
RT flavoproteins.";
RL Appl. Environ. Microbiol. 70:3566-3574(2004).
RN [5]
RP FUNCTION, AND DISRUPTION PHENOTYPE.
RC STRAIN=K12 / AB1157;
RX PubMed=17504490; DOI=10.1111/j.1462-2920.2007.01272.x;
RA Gonzalez-Perez M.M., van Dillewijn P., Wittich R.M., Ramos J.L.;
RT "Escherichia coli has multiple enzymes that attack TNT and release nitrogen
RT for growth.";
RL Environ. Microbiol. 9:1535-1540(2007).
RN [6]
RP INDUCTION.
RC STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX PubMed=18567656; DOI=10.1128/jb.00459-08;
RA Umezawa Y., Shimada T., Kori A., Yamada K., Ishihama A.;
RT "The uncharacterized transcription factor YdhM is the regulator of the nemA
RT gene, encoding N-ethylmaleimide reductase.";
RL J. Bacteriol. 190:5890-5897(2008).
RN [7]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND INDUCTION.
RC STRAIN=K12;
RX PubMed=23506073; DOI=10.1111/mmi.12192;
RA Lee C., Shin J., Park C.;
RT "Novel regulatory system nemRA-gloA for electrophile reduction in
RT Escherichia coli K-12.";
RL Mol. Microbiol. 88:395-412(2013).
RN [8]
RP FUNCTION, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=23527133; DOI=10.1371/journal.pone.0059200;
RA Robins K.J., Hooks D.O., Rehm B.H., Ackerley D.F.;
RT "Escherichia coli NemA is an efficient chromate reductase that can be
RT biologically immobilized to provide a cell free system for remediation of
RT hexavalent chromium.";
RL PLoS ONE 8:E59200-E59200(2013).
RN [9]
RP FUNCTION, INDUCTION, AND DISRUPTION PHENOTYPE.
RX PubMed=23536188; DOI=10.1074/jbc.m113.454421;
RA Gray M.J., Wholey W.Y., Parker B.W., Kim M., Jakob U.;
RT "NemR is a bleach-sensing transcription factor.";
RL J. Biol. Chem. 288:13789-13798(2013).
RN [10]
RP INDUCTION.
RX PubMed=23646895; DOI=10.1111/mmi.12234;
RA Ozyamak E., de Almeida C., de Moura A.P., Miller S., Booth I.R.;
RT "Integrated stress response of Escherichia coli to methylglyoxal:
RT transcriptional readthrough from the nemRA operon enhances protection
RT through increased expression of glyoxalase I.";
RL Mol. Microbiol. 88:936-950(2013).
CC -!- FUNCTION: Involved in the degradation of toxic compounds
CC (PubMed:9013822, PubMed:15184158, PubMed:17504490, PubMed:23527133,
CC PubMed:23506073, PubMed:23536188). Can use a variety of substrates,
CC including the nitrate ester explosives glycerol trinitrate (GTN) and
CC pentaerythritol tetranitrate (PETN), chromate and various electrophiles
CC such as quinones (PubMed:15184158, PubMed:23527133, PubMed:23506073).
CC Involved in resistance to hypochlorous acid (HOCl), which is the active
CC component of household bleach and a powerful antimicrobial during the
CC innate immune response (PubMed:23536188). Catalyzes the reduction of N-
CC ethylmaleimide (NEM) to N-ethylsuccinimide (PubMed:9013822,
CC PubMed:23506073). Together with NfsA and NfsB, can use the
CC nitroaromatic explosive 2,4,6-trinitrotoluene (TNT) (PubMed:17504490).
CC {ECO:0000269|PubMed:15184158, ECO:0000269|PubMed:17504490,
CC ECO:0000269|PubMed:23506073, ECO:0000269|PubMed:23527133,
CC ECO:0000269|PubMed:23536188, ECO:0000269|PubMed:9013822}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H(+) + N-ethylmaleimide + NADPH = N-ethylsuccinimide +
CC NADP(+); Xref=Rhea:RHEA:35523, ChEBI:CHEBI:15378, ChEBI:CHEBI:44348,
CC ChEBI:CHEBI:44485, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349;
CC Evidence={ECO:0000269|PubMed:23506073, ECO:0000269|PubMed:9013822};
CC -!- COFACTOR:
CC Name=FMN; Xref=ChEBI:CHEBI:58210;
CC Evidence={ECO:0000250|UniProtKB:P42593};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=27.13 uM for NEM {ECO:0000269|PubMed:23506073};
CC KM=23 uM for chromate (in the presence of NADPH)
CC {ECO:0000269|PubMed:23527133};
CC KM=16 uM for chromate (in the presence of NADH)
CC {ECO:0000269|PubMed:23527133};
CC KM=0.29 mM for menadione {ECO:0000269|PubMed:23506073};
CC KM=0.31 mM for ubiquinone 0 {ECO:0000269|PubMed:23506073};
CC KM=0.78 mM for ubiquinone 10 {ECO:0000269|PubMed:23506073};
CC KM=6.43 mM for acrolein {ECO:0000269|PubMed:23506073};
CC KM=11.52 mM for menaquinone {ECO:0000269|PubMed:23506073};
CC KM=25.63 mM for methylglyoxal {ECO:0000269|PubMed:23506073};
CC KM=318 mM for glyoxal {ECO:0000269|PubMed:23506073};
CC KM=480 mM for glycolaldehyde {ECO:0000269|PubMed:23506073};
CC Note=kcat is 354000 sec(-1) with NEM as substrate. kcat is 123000
CC sec(-1) with ubiquinone 0 as substrate. kcat is 14200 sec(-1) with
CC menadione as substrate (PubMed:23506073). kcat is 2.1 sec(-1) with
CC chromate as substrate in the presence of NADPH. kcat is 1 sec(-1)
CC with chromate as substrate in the presence of NADH (PubMed:23527133).
CC {ECO:0000269|PubMed:23506073, ECO:0000269|PubMed:23527133};
CC -!- INDUCTION: Repressed by NemR. Induced by N-ethylmaleimide and reactive
CC electrophilic species (RES) such as quinones, glyoxals and
CC methylglyoxal (PubMed:18567656, PubMed:23506073, PubMed:23646895). Up-
CC regulated by HOCl (PubMed:23536188). {ECO:0000269|PubMed:18567656,
CC ECO:0000269|PubMed:23506073, ECO:0000269|PubMed:23536188,
CC ECO:0000269|PubMed:23646895}.
CC -!- DISRUPTION PHENOTYPE: Mutant is still able to grow and use TNT as the
CC sole nitrogen source (PubMed:17504490). Deletion of the gene increases
CC the HOCl sensitivity. Mutant shows no defect in survival of
CC methylglyoxal stress (PubMed:23536188). {ECO:0000269|PubMed:17504490,
CC ECO:0000269|PubMed:23536188}.
CC -!- SIMILARITY: Belongs to the NADH:flavin oxidoreductase/NADH oxidase
CC family. {ECO:0000305}.
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DR EMBL; D86931; BAA13186.1; -; Genomic_DNA.
DR EMBL; U00096; AAC74722.1; -; Genomic_DNA.
DR EMBL; AP009048; BAE76493.1; -; Genomic_DNA.
DR PIR; JC5605; JC5605.
DR RefSeq; NP_416167.1; NC_000913.3.
DR RefSeq; WP_000093589.1; NZ_SSZK01000001.1.
DR AlphaFoldDB; P77258; -.
DR SMR; P77258; -.
DR BioGRID; 4259388; 46.
DR IntAct; P77258; 3.
DR STRING; 511145.b1650; -.
DR SWISS-2DPAGE; P77258; -.
DR jPOST; P77258; -.
DR PaxDb; P77258; -.
DR PRIDE; P77258; -.
DR EnsemblBacteria; AAC74722; AAC74722; b1650.
DR EnsemblBacteria; BAE76493; BAE76493; BAE76493.
DR GeneID; 946164; -.
DR KEGG; ecj:JW1642; -.
DR KEGG; eco:b1650; -.
DR PATRIC; fig|1411691.4.peg.609; -.
DR EchoBASE; EB3316; -.
DR eggNOG; COG1902; Bacteria.
DR HOGENOM; CLU_012153_0_2_6; -.
DR InParanoid; P77258; -.
DR OMA; MQIMHGG; -.
DR PhylomeDB; P77258; -.
DR BioCyc; EcoCyc:G6890-MON; -.
DR BioCyc; MetaCyc:G6890-MON; -.
DR PRO; PR:P77258; -.
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:0008748; F:N-ethylmaleimide reductase activity; IDA:EcoCyc.
DR GO; GO:0046857; F:oxidoreductase activity, acting on other nitrogenous compounds as donors, with NAD or NADP as acceptor; IDA:EcoCyc.
DR GO; GO:0046256; P:2,4,6-trinitrotoluene catabolic process; IMP:EcoCyc.
DR GO; GO:0018937; P:nitroglycerin metabolic process; IDA:EcoCyc.
DR GO; GO:0006805; P:xenobiotic metabolic process; IDA:EcoCyc.
DR Gene3D; 3.20.20.70; -; 1.
DR InterPro; IPR013785; Aldolase_TIM.
DR InterPro; IPR001155; OxRdtase_FMN_N.
DR InterPro; IPR045247; Oye-like.
DR PANTHER; PTHR22893; PTHR22893; 1.
DR Pfam; PF00724; Oxidored_FMN; 1.
PE 1: Evidence at protein level;
KW Flavoprotein; FMN; NADP; Oxidoreductase; Reference proteome.
FT CHAIN 1..365
FT /note="N-ethylmaleimide reductase"
FT /id="PRO_0000194481"
FT ACT_SITE 187
FT /note="Proton donor"
FT /evidence="ECO:0000250|UniProtKB:P42593"
FT BINDING 101
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000250|UniProtKB:P42593"
FT BINDING 234
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000250|UniProtKB:P42593"
FT BINDING 324..325
FT /ligand="FMN"
FT /ligand_id="ChEBI:CHEBI:58210"
FT /evidence="ECO:0000250|UniProtKB:P42593"
SQ SEQUENCE 365 AA; 39516 MW; EFCFE04B42CE9C7C CRC64;
MSSEKLYSPL KVGAITAANR IFMAPLTRLR SIEPGDIPTP LMAEYYRQRA SAGLIISEAT
QISAQAKGYA GAPGIHSPEQ IAAWKKITAG VHAENGHMAV QLWHTGRISH ASLQPGGQAP
VAPSALSAGT RTSLRDENGQ AIRVETSMPR ALELEEIPGI VNDFRQAIAN AREAGFDLVE
LHSAHGYLLH QFLSPSSNHR TDQYGGSVEN RARLVLEVVD AGIEEWGADR IGIRVSPIGT
FQNTDNGPNE EADALYLIEQ LGKRGIAYLH MSEPDWAGGE PYTDAFREKV RARFHGPIIG
AGAYTVEKAE TLIGKGLIDA VAFGRDWIAN PDLVARLQRK AELNPQRAES FYGGGAEGYT
DYPTL
