CGR2_EGGLE
ID CGR2_EGGLE Reviewed; 560 AA.
AC C8WLM1;
DT 22-APR-2020, integrated into UniProtKB/Swiss-Prot.
DT 03-NOV-2009, sequence version 1.
DT 03-AUG-2022, entry version 60.
DE RecName: Full=Digoxin reductase {ECO:0000305|PubMed:29761785};
DE EC=1.3.2.- {ECO:0000269|PubMed:29761785};
DE AltName: Full=Cardenolide reductase {ECO:0000305|PubMed:29761785};
DE AltName: Full=Cardiac glycoside reductase operon protein 2 {ECO:0000303|PubMed:23869020};
DE Flags: Precursor;
GN Name=cgr2 {ECO:0000303|PubMed:23869020};
GN OrderedLocusNames=Elen_2529 {ECO:0000312|EMBL:ACV56484.1};
OS Eggerthella lenta (strain ATCC 25559 / DSM 2243 / CCUG 17323 / JCM 9979 /
OS KCTC 3265 / NCTC 11813 / VPI 0255 / 1899 B) (Eubacterium lentum).
OC Bacteria; Actinobacteria; Coriobacteriia; Eggerthellales; Eggerthellaceae;
OC Eggerthella.
OX NCBI_TaxID=479437;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 25559 / DSM 2243 / CCUG 17323 / JCM 9979 / KCTC 3265 / NCTC
RC 11813 / VPI 0255 / 1899 B;
RX PubMed=21304654; DOI=10.4056/sigs.33592;
RA Saunders E., Pukall R., Abt B., Lapidus A., Glavina Del Rio T.,
RA Copeland A., Tice H., Cheng J.F., Lucas S., Chen F., Nolan M., Bruce D.,
RA Goodwin L., Pitluck S., Ivanova N., Mavromatis K., Ovchinnikova G.,
RA Pati A., Chen A., Palaniappan K., Land M., Hauser L., Chang Y.J.,
RA Jeffries C.D., Chain P., Meincke L., Sims D., Brettin T., Detter J.C.,
RA Goker M., Bristow J., Eisen J.A., Markowitz V., Hugenholtz P.,
RA Kyrpides N.C., Klenk H.P., Han C.;
RT "Complete genome sequence of Eggerthella lenta type strain (IPP VPI
RT 0255).";
RL Stand. Genomic Sci. 1:174-182(2009).
RN [2]
RP INDUCTION.
RC STRAIN=ATCC 25559 / DSM 2243 / CCUG 17323 / JCM 9979 / KCTC 3265 / NCTC
RC 11813 / VPI 0255 / 1899 B;
RX PubMed=23869020; DOI=10.1126/science.1235872;
RA Haiser H.J., Gootenberg D.B., Chatman K., Sirasani G., Balskus E.P.,
RA Turnbaugh P.J.;
RT "Predicting and manipulating cardiac drug inactivation by the human gut
RT bacterium Eggerthella lenta.";
RL Science 341:295-298(2013).
RN [3]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBSTRATE SPECIFICITY, MUTAGENESIS OF CYS-82; CYS-158; CYS-187; CYS-265;
RP CYS-327; TYR-532; CYS-535 AND GLY-536, REACTION MECHANISM, SUBUNIT, AND
RP SUBCELLULAR LOCATION.
RC STRAIN=ATCC 25559 / DSM 2243 / CCUG 17323 / JCM 9979 / KCTC 3265 / NCTC
RC 11813 / VPI 0255 / 1899 B;
RX PubMed=29761785; DOI=10.7554/elife.33953;
RA Koppel N., Bisanz J.E., Pandelia M.E., Turnbaugh P.J., Balskus E.P.;
RT "Discovery and characterization of a prevalent human gut bacterial enzyme
RT sufficient for the inactivation of a family of plant toxins.";
RL Elife 7:E33953-E33953(2018).
CC -!- FUNCTION: Involved in the inactivation of the cardiac medication and
CC plant natural product digoxin, thus decreasing drug efficacy and
CC toxicity. Catalyzes the reduction of the alpha,beta-unsaturated
CC butyrolactone ring of digoxin to the inactive metabolite
CC dihydrodigoxin. Likely uses the cytochrome Cgr1 as the physiological
CC electron donor, encoded by the adjacent gene in the locus. Only reduces
CC digoxin and other cardenolide toxins, such as digitoxin, digoxigenin,
CC ouabain and ouabagenin. Therefore is a specialized enzyme present in
CC some gut bacteria E.lenta that protects their human host against
CC ingested plant toxins. {ECO:0000269|PubMed:29761785}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=digoxin + 2 Fe(II)-[cytochrome c] + 3 H(+) = dihydrodigoxin +
CC 2 Fe(III)-[cytochrome c]; Xref=Rhea:RHEA:62528, Rhea:RHEA-COMP:10350,
CC Rhea:RHEA-COMP:14399, ChEBI:CHEBI:15378, ChEBI:CHEBI:29033,
CC ChEBI:CHEBI:29034, ChEBI:CHEBI:71002, ChEBI:CHEBI:145795;
CC Evidence={ECO:0000269|PubMed:29761785, ECO:0000305};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62529;
CC Evidence={ECO:0000305|PubMed:29761785};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=digitoxin + 2 Fe(II)-[cytochrome c] + 3 H(+) =
CC dihydrodigitoxin + 2 Fe(III)-[cytochrome c]; Xref=Rhea:RHEA:62532,
CC Rhea:RHEA-COMP:10350, Rhea:RHEA-COMP:14399, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29033, ChEBI:CHEBI:29034, ChEBI:CHEBI:145796,
CC ChEBI:CHEBI:282234; Evidence={ECO:0000269|PubMed:29761785,
CC ECO:0000305};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62533;
CC Evidence={ECO:0000305|PubMed:29761785};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=digoxigenin + 2 Fe(II)-[cytochrome c] + 3 H(+) =
CC dihydrodigoxigenin + 2 Fe(III)-[cytochrome c]; Xref=Rhea:RHEA:62536,
CC Rhea:RHEA-COMP:10350, Rhea:RHEA-COMP:14399, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29033, ChEBI:CHEBI:29034, ChEBI:CHEBI:71004,
CC ChEBI:CHEBI:145797; Evidence={ECO:0000269|PubMed:29761785,
CC ECO:0000305};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62537;
CC Evidence={ECO:0000305|PubMed:29761785};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 Fe(II)-[cytochrome c] + 3 H(+) + ouabain = dihydroouabain +
CC 2 Fe(III)-[cytochrome c]; Xref=Rhea:RHEA:62540, Rhea:RHEA-COMP:10350,
CC Rhea:RHEA-COMP:14399, ChEBI:CHEBI:15378, ChEBI:CHEBI:29033,
CC ChEBI:CHEBI:29034, ChEBI:CHEBI:131146, ChEBI:CHEBI:145798;
CC Evidence={ECO:0000269|PubMed:29761785, ECO:0000305};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62541;
CC Evidence={ECO:0000305|PubMed:29761785};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=2 Fe(II)-[cytochrome c] + 3 H(+) + ouabagenin =
CC dihydroouabagenin + 2 Fe(III)-[cytochrome c]; Xref=Rhea:RHEA:62544,
CC Rhea:RHEA-COMP:10350, Rhea:RHEA-COMP:14399, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:29033, ChEBI:CHEBI:29034, ChEBI:CHEBI:145789,
CC ChEBI:CHEBI:145799; Evidence={ECO:0000269|PubMed:29761785,
CC ECO:0000305};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:62545;
CC Evidence={ECO:0000305|PubMed:29761785};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:29761785};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000269|PubMed:29761785};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=94.6 uM for digoxin {ECO:0000269|PubMed:29761785};
CC Note=kcat is 0.23 sec(-1) for the reduction of digoxin.
CC {ECO:0000269|PubMed:29761785};
CC -!- SUBUNIT: May form a membrane-associated complex with Cgr1.
CC {ECO:0000305|PubMed:29761785}.
CC -!- SUBCELLULAR LOCATION: Cell membrane; Peripheral membrane protein;
CC Extracellular side {ECO:0000305|PubMed:29761785}.
CC -!- INDUCTION: Is highly up-regulated by digoxin and other cardiac
CC glycosides containing unsaturated butyrolactone rings, such as
CC digitoxin, digoxigenin, and, to a lesser extent, ouabain. Is repressed
CC by arginine but not by ornithine. Part of an operon that consists of
CC cgr1 and cgr2. {ECO:0000269|PubMed:23869020}.
CC -!- PTM: Predicted to be exported by the Tat system. The position of the
CC signal peptide cleavage has not been experimentally proven.
CC {ECO:0000255|PROSITE-ProRule:PRU00648}.
CC -!- MISCELLANEOUS: Digoxin is a toxic chemical produced by plants that, in
CC low doses, can be used to treat heart failure and arrhythmia. cgr
CC operon presence and abundance predicted the extent of drug inactivation
CC by human gut microbial communities in ex vivo incubations, and is
CC restricted to E.lenta species. Data about the gut microbes in nearly
CC 1,900 people from three continents revealed that bacteria that can
CC produce Cgr2 were present in the guts of more than 40% of the
CC individuals, although often in low abundance.
CC {ECO:0000269|PubMed:23869020, ECO:0000269|PubMed:29761785}.
CC -!- SIMILARITY: Belongs to the FAD-dependent oxidoreductase 2 family.
CC {ECO:0000305}.
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DR EMBL; CP001726; ACV56484.1; -; Genomic_DNA.
DR RefSeq; WP_015761237.1; NC_013204.1.
DR AlphaFoldDB; C8WLM1; -.
DR SMR; C8WLM1; -.
DR STRING; 479437.Elen_2529; -.
DR EnsemblBacteria; ACV56484; ACV56484; Elen_2529.
DR KEGG; ele:Elen_2529; -.
DR eggNOG; COG1053; Bacteria.
DR HOGENOM; CLU_011398_4_3_11; -.
DR OMA; TRYFRED; -.
DR OrthoDB; 153138at2; -.
DR BioCyc; ELEN479437:G1GFY-2552-MON; -.
DR Proteomes; UP000001377; Chromosome.
DR GO; GO:0005886; C:plasma membrane; IEA:UniProtKB-SubCell.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0033765; F:steroid dehydrogenase activity, acting on the CH-CH group of donors; IEA:UniProt.
DR GO; GO:0009636; P:response to toxic substance; IEA:UniProtKB-KW.
DR GO; GO:0008202; P:steroid metabolic process; IEA:UniProt.
DR Gene3D; 3.50.50.60; -; 1.
DR Gene3D; 3.90.700.10; -; 1.
DR InterPro; IPR003953; FAD-binding_2.
DR InterPro; IPR036188; FAD/NAD-bd_sf.
DR InterPro; IPR027477; Succ_DH/fumarate_Rdtase_cat_sf.
DR InterPro; IPR006311; TAT_signal.
DR InterPro; IPR019546; TAT_signal_bac_arc.
DR Pfam; PF00890; FAD_binding_2; 1.
DR SUPFAM; SSF51905; SSF51905; 1.
DR SUPFAM; SSF56425; SSF56425; 1.
DR TIGRFAMs; TIGR01409; TAT_signal_seq; 1.
DR PROSITE; PS51318; TAT; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Cell membrane; Detoxification; FAD; Flavoprotein; Iron;
KW Iron-sulfur; Membrane; Metal-binding; Oxidoreductase; Reference proteome;
KW Signal.
FT SIGNAL 1..48
FT /note="Tat-type signal"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00648"
FT CHAIN 49..560
FT /note="Digoxin reductase"
FT /id="PRO_5002992288"
FT MUTAGEN 82
FT /note="C->A: Decrease in catalytic activity. Still binds a
FT [4Fe-4S] cluster."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 158
FT /note="C->A: Decrease in catalytic activity. Still binds a
FT [4Fe-4S] cluster."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 187
FT /note="C->A: Decrease in catalytic activity. Still binds a
FT [4Fe-4S] cluster."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 265
FT /note="C->A: Decrease in catalytic activity. Still binds a
FT [4Fe-4S] cluster."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 327
FT /note="C->A: Decrease in catalytic activity. Still binds a
FT [4Fe-4S] cluster."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 532
FT /note="Y->F: No effect on catalytic activity."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 535
FT /note="C->A: Decrease in catalytic activity. Still binds a
FT [4Fe-4S] cluster."
FT /evidence="ECO:0000269|PubMed:29761785"
FT MUTAGEN 536
FT /note="G->A: Decrease in catalytic activity."
FT /evidence="ECO:0000269|PubMed:29761785"
SQ SEQUENCE 560 AA; 59161 MW; 102A4A8E0A3FA621 CRC64;
MEYGKCRGIE RGMGRRDFLK AATLLGATAA GAGMLAGCAP KSASEAQAQT APAATGGLDP
ADVDWKYETD VVIVGSGSGG TCAAIEAAEA GADVVVFEKD KAMYGGNSAL CGGYMLAAGW
STQEEITGYA GDTGEAFANQ MLRWSQGLGN QDMIREACLR SGEAVDWMMD TGRTYEGASP
LPPVWSCGDT EADVVPRSVY NHNAYGATEG HMATLKKRAE SLSNIEIEMG CEVAHILKNA
EGSVIGVQLA DGSFAKARKG VVMACASVDN NLEMSKDLGL MQNVWGLTLE GAGLLAPGNP
DMDSNTGDGV RMLREIGAEL CMQQAVCMND SIYVGGISDW GMSEILGKDV NIHDSSNIDA
ILVDKTGRRF CQDDAEWGYV MHECAQAAWK QGFTPDDPTT GYIFYVYDAT GAPFFEMKGH
TPDTCDTTFS ADSVDGLAEF IGCDPTALAS EVERWNSFCE AGLDADFGRR ANMAPIATPP
FYCDVVRPGP MGTFAGAKSN VEAEIIGLDG NPIPRLYGAG CIIGGNVSGA FYFGCGWSIT
NTVVWGREAG RNVAALEPWE
