ETHA_MYCTU
ID ETHA_MYCTU Reviewed; 489 AA.
AC P9WNF9; L0TDR6; P96223; Q7D4Q8;
DT 16-APR-2014, integrated into UniProtKB/Swiss-Prot.
DT 16-APR-2014, sequence version 1.
DT 03-AUG-2022, entry version 43.
DE RecName: Full=FAD-containing monooxygenase EthA {ECO:0000303|PubMed:11823459};
DE EC=1.14.13.- {ECO:0000269|PubMed:14610090};
DE AltName: Full=Baeyer-Villiger monooxygenase EtaA {ECO:0000303|PubMed:14610090};
DE Short=BVMO {ECO:0000303|PubMed:14610090};
DE AltName: Full=Prodrug activator EtaA {ECO:0000303|PubMed:14610090};
GN Name=ethA {ECO:0000303|PubMed:10869356};
GN Synonyms=etaA {ECO:0000303|PubMed:10944230, ECO:0000303|PubMed:11823459};
GN OrderedLocusNames=Rv3854c;
OS Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv).
OC Bacteria; Actinobacteria; Corynebacteriales; Mycobacteriaceae;
OC Mycobacterium; Mycobacterium tuberculosis complex.
OX NCBI_TaxID=83332;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=9634230; DOI=10.1038/31159;
RA Cole S.T., Brosch R., Parkhill J., Garnier T., Churcher C.M., Harris D.E.,
RA Gordon S.V., Eiglmeier K., Gas S., Barry C.E. III, Tekaia F., Badcock K.,
RA Basham D., Brown D., Chillingworth T., Connor R., Davies R.M., Devlin K.,
RA Feltwell T., Gentles S., Hamlin N., Holroyd S., Hornsby T., Jagels K.,
RA Krogh A., McLean J., Moule S., Murphy L.D., Oliver S., Osborne J.,
RA Quail M.A., Rajandream M.A., Rogers J., Rutter S., Seeger K., Skelton S.,
RA Squares S., Squares R., Sulston J.E., Taylor K., Whitehead S.,
RA Barrell B.G.;
RT "Deciphering the biology of Mycobacterium tuberculosis from the complete
RT genome sequence.";
RL Nature 393:537-544(1998).
RN [2]
RP FUNCTION AS AN ACTIVATOR OF ETHIONAMIDE, AND TRANSCRIPTIONAL REGULATION.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=10869356; DOI=10.1074/jbc.m003744200;
RA Baulard A.R., Betts J.C., Engohang-Ndong J., Quan S., McAdam R.A.,
RA Brennan P.J., Locht C., Besra G.S.;
RT "Activation of the pro-drug ethionamide is regulated in mycobacteria.";
RL J. Biol. Chem. 275:28326-28331(2000).
RN [3]
RP FUNCTION AS AN ACTIVATOR OF ETHIONAMIDE, AND DISRUPTION PHENOTYPE.
RX PubMed=10944230; DOI=10.1073/pnas.97.17.9677;
RA DeBarber A.E., Mdluli K., Bosman M., Bekker L.G., Barry C.E. III;
RT "Ethionamide activation and sensitivity in multidrug-resistant
RT Mycobacterium tuberculosis.";
RL Proc. Natl. Acad. Sci. U.S.A. 97:9677-9682(2000).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR,
RP SUBSTRATE SPECIFICITY, AND SUBCELLULAR LOCATION.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=11823459; DOI=10.1074/jbc.m110751200;
RA Vannelli T.A., Dykman A., Ortiz de Montellano P.R.;
RT "The antituberculosis drug ethionamide is activated by a flavoprotein
RT monooxygenase.";
RL J. Biol. Chem. 277:12824-12829(2002).
RN [5]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE SPECIFICITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, AND SUBUNIT.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=14610090; DOI=10.1074/jbc.m307770200;
RA Fraaije M.W., Kamerbeek N.M., Heidekamp A.J., Fortin R., Janssen D.B.;
RT "The prodrug activator EtaA from Mycobacterium tuberculosis is a Baeyer-
RT Villiger monooxygenase.";
RL J. Biol. Chem. 279:3354-3360(2004).
RN [6]
RP MUTAGENESIS OF TYR-84.
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=19412174; DOI=10.1038/nm.1950;
RA Willand N., Dirie B., Carette X., Bifani P., Singhal A., Desroses M.,
RA Leroux F., Willery E., Mathys V., Deprez-Poulain R., Delcroix G.,
RA Frenois F., Aumercier M., Locht C., Villeret V., Deprez B., Baulard A.R.;
RT "Synthetic EthR inhibitors boost antituberculous activity of ethionamide.";
RL Nat. Med. 15:537-544(2009).
RN [7]
RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS].
RC STRAIN=ATCC 25618 / H37Rv;
RX PubMed=21969609; DOI=10.1074/mcp.m111.011627;
RA Kelkar D.S., Kumar D., Kumar P., Balakrishnan L., Muthusamy B., Yadav A.K.,
RA Shrivastava P., Marimuthu A., Anand S., Sundaram H., Kingsbury R.,
RA Harsha H.C., Nair B., Prasad T.S., Chauhan D.S., Katoch K., Katoch V.M.,
RA Kumar P., Chaerkady R., Ramachandran S., Dash D., Pandey A.;
RT "Proteogenomic analysis of Mycobacterium tuberculosis by high resolution
RT mass spectrometry.";
RL Mol. Cell. Proteomics 10:M111.011627-M111.011627(2011).
CC -!- FUNCTION: Monooxygenase able to convert a wide range of ketones to the
CC corresponding esters or lactones via a Baeyer-Villiger oxidation
CC reaction. Can act on long-chain aliphatic ketones (2-hexanone to 2-
CC dodecanone) and on aromatic ketones (phenylacetone and benzylacetone).
CC Is also able to catalyze enantioselective sulfoxidation of methyl-p-
CC tolylsulfide. In vivo, likely functions as a BVMO, but the exact nature
CC of the physiological substrate(s) remains to be established.
CC {ECO:0000269|PubMed:14610090}.
CC -!- FUNCTION: Is responsible for the activation of several thiocarbamide-
CC containing pro-drugs into cytotoxic species. Thus, catalyzes the
CC oxidation of the antitubercular pro-drug ethionamide (ETH) to the
CC corresponding sulfoxide, which is further oxidized by EthA to 2-ethyl-
CC 4-amidopyridine, presumably via the unstable doubly oxidized sulfinic
CC acid intermediate; the final metabolite 2-ethyl-4-amidopyridine has no
CC antitubercular activity, so the cytotoxic species is a metabolite
CC intermediate formed by EthA. Also oxidizes thiacetazone (TAC),
CC thiobenzamide, and isothionicotinamide and therefore is probably
CC responsible, as suggested by the observation of crossover resistance,
CC for the oxidative activation of these other thioamide antitubercular
CC drugs. {ECO:0000269|PubMed:10869356, ECO:0000269|PubMed:10944230,
CC ECO:0000269|PubMed:11823459, ECO:0000269|PubMed:14610090}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ethionamide + H(+) + NADPH + O2 = ethionamide S-oxide + H2O +
CC NADP(+); Xref=Rhea:RHEA:47616, ChEBI:CHEBI:4885, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:15379, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349, ChEBI:CHEBI:87805;
CC Evidence={ECO:0000269|PubMed:11823459, ECO:0000269|PubMed:14610090};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:11823459, ECO:0000269|PubMed:14610090};
CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:11823459,
CC ECO:0000269|PubMed:14610090};
CC -!- ACTIVITY REGULATION: Activity can be increased by one order of
CC magnitude by adding bovine serum albumin in vitro. This result suggests
CC that the enzyme is activated in vivo by protein-protein or interactions
CC with other cellular components. {ECO:0000269|PubMed:14610090}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=194 uM for ethionamide {ECO:0000269|PubMed:11823459};
CC KM=340 uM for ethionamide {ECO:0000269|PubMed:14610090};
CC KM=10 uM for NADPH {ECO:0000269|PubMed:14610090};
CC KM=61 uM for phenylacetone {ECO:0000269|PubMed:14610090};
CC KM=520 uM for benzylacetone {ECO:0000269|PubMed:14610090};
CC KM=200 uM for 2-dodecanone {ECO:0000269|PubMed:14610090};
CC Note=kcat is 0.027 sec(-1) with ethionamide as substrate. kcat is
CC 0.017 sec(-1) with phenylacetone as substrate. kcat is 0.021 sec(-1)
CC with benzylacetone as substrate. kcat is 0.023 sec(-1) with 2-
CC dodecanone as substrate. {ECO:0000269|PubMed:14610090};
CC -!- SUBUNIT: Exists as a mixture of relatively large homooligomers ranging
CC from 200 to 600 kDa. {ECO:0000269|PubMed:14610090}.
CC -!- SUBCELLULAR LOCATION: Cell membrane {ECO:0000303|PubMed:11823459}.
CC Note=Is most likely membrane-associated. {ECO:0000303|PubMed:11823459}.
CC -!- INDUCTION: Repressed by the transcriptional regulator EthR.
CC {ECO:0000269|PubMed:10869356}.
CC -!- DISRUPTION PHENOTYPE: Inactivation of this gene leads to a strong
CC resistance to ETH. {ECO:0000269|PubMed:10944230}.
CC -!- SIMILARITY: Belongs to the FAD-binding monooxygenase family.
CC {ECO:0000305}.
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DR EMBL; AL123456; CCP46683.1; -; Genomic_DNA.
DR PIR; C70655; C70655.
DR RefSeq; NP_218371.1; NC_000962.3.
DR RefSeq; WP_003899731.1; NZ_NVQJ01000057.1.
DR AlphaFoldDB; P9WNF9; -.
DR SMR; P9WNF9; -.
DR STRING; 83332.Rv3854c; -.
DR PaxDb; P9WNF9; -.
DR DNASU; 886175; -.
DR GeneID; 886175; -.
DR KEGG; mtu:Rv3854c; -.
DR TubercuList; Rv3854c; -.
DR eggNOG; COG2072; Bacteria.
DR OMA; ASWTLKC; -.
DR PhylomeDB; P9WNF9; -.
DR BioCyc; MetaCyc:G185E-8152-MON; -.
DR SABIO-RK; P9WNF9; -.
DR Proteomes; UP000001584; Chromosome.
DR GO; GO:0005829; C:cytosol; HDA:MTBBASE.
DR GO; GO:0009274; C:peptidoglycan-based cell wall; HDA:UniProtKB.
DR GO; GO:0005886; C:plasma membrane; IDA:MTBBASE.
DR GO; GO:0071949; F:FAD binding; IDA:UniProtKB.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IDA:UniProtKB.
DR GO; GO:0004499; F:N,N-dimethylaniline monooxygenase activity; IDA:MTBBASE.
DR GO; GO:0070402; F:NADPH binding; IDA:UniProtKB.
DR GO; GO:0016709; F:oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen, NAD(P)H as one donor, and incorporation of one atom of oxygen; IDA:UniProtKB.
DR GO; GO:0033776; F:phenylacetone monooxygenase activity; IDA:UniProtKB.
DR GO; GO:0006805; P:xenobiotic metabolic process; IDA:UniProtKB.
DR Gene3D; 3.50.50.60; -; 3.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR020946; Flavin_mOase-like.
DR Pfam; PF00743; FMO-like; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
PE 1: Evidence at protein level;
KW Cell membrane; FAD; Flavoprotein; Membrane; Monooxygenase; NADP;
KW Oxidoreductase; Reference proteome.
FT CHAIN 1..489
FT /note="FAD-containing monooxygenase EthA"
FT /id="PRO_0000398581"
FT BINDING 15
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 36
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 44..47
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 54..56
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 56
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 104
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 183..189
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT BINDING 207..208
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT SITE 292
FT /note="Transition state stabilizer"
FT /evidence="ECO:0000250|UniProtKB:Q47PU3"
FT MUTAGEN 84
FT /note="Y->C: ETH-resistant."
FT /evidence="ECO:0000269|PubMed:19412174"
SQ SEQUENCE 489 AA; 55326 MW; 844611B7E831180D CRC64;
MTEHLDVVIV GAGISGVSAA WHLQDRCPTK SYAILEKRES MGGTWDLFRY PGIRSDSDMY
TLGFRFRPWT GRQAIADGKP ILEYVKSTAA MYGIDRHIRF HHKVISADWS TAENRWTVHI
QSHGTLSALT CEFLFLCSGY YNYDEGYSPR FAGSEDFVGP IIHPQHWPED LDYDAKNIVV
IGSGATAVTL VPALADSGAK HVTMLQRSPT YIVSQPDRDG IAEKLNRWLP ETMAYTAVRW
KNVLRQAAVY SACQKWPRRM RKMFLSLIQR QLPEGYDVRK HFGPHYNPWD QRLCLVPNGD
LFRAIRHGKV EVVTDTIERF TATGIRLNSG RELPADIIIT ATGLNLQLFG GATATIDGQQ
VDITTTMAYK GMMLSGIPNM AYTVGYTNAS WTLKADLVSE FVCRLLNYMD DNGFDTVVVE
RPGSDVEERP FMEFTPGYVL RSLDELPKQG SRTPWRLNQN YLRDIRLIRR GKIDDEGLRF
AKRPAPVGV
