MDFA_ECOLI
ID MDFA_ECOLI Reviewed; 410 AA.
AC P0AEY8; P71226; P75807; Q46966;
DT 20-DEC-2005, integrated into UniProtKB/Swiss-Prot.
DT 20-DEC-2005, sequence version 1.
DT 03-AUG-2022, entry version 125.
DE RecName: Full=Multidrug transporter MdfA;
DE AltName: Full=Chloramphenicol resistance pump Cmr;
GN Name=mdfA; Synonyms=cmlA, cmr; OrderedLocusNames=b0842, JW0826;
OS Escherichia coli (strain K12).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=83333;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=K12;
RX PubMed=8655497; DOI=10.1128/jb.178.11.3188-3193.1996;
RA Nilsen I.W., Bakke I., Vader A., Olsvik O., El-Gewely M.R.;
RT "Isolation of cmr, a novel Escherichia coli chloramphenicol resistance gene
RT encoding a putative efflux pump.";
RL J. Bacteriol. 178:3188-3193(1996).
RN [2]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND FUNCTION.
RC STRAIN=ATCC 33694 / HB101;
RX PubMed=9079913; DOI=10.1128/jb.179.7.2274-2280.1997;
RA Edgar R., Bibi E.;
RT "MdfA, an Escherichia coli multidrug resistance protein with an
RT extraordinarily broad spectrum of drug recognition.";
RL J. Bacteriol. 179:2274-2280(1997).
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 FUNCTION.
RC STRAIN=K12 / C600 / CR34 / ATCC 23724 / DSM 3925 / LMG 3041 / NCIB 10222;
RX PubMed=9811673; DOI=10.1128/jb.180.22.6072-6075.1998;
RA Bohn C., Bouloc P.;
RT "The Escherichia coli cmlA gene encodes the multidrug efflux pump Cmr/MdfA
RT and is responsible for isopropyl-beta-D-thiogalactopyranoside exclusion and
RT spectinomycin sensitivity.";
RL J. Bacteriol. 180:6072-6075(1998).
RN [7]
RP FUNCTION AS AN ANTIPORTER, ACTIVITY REGULATION, AND BIOPHYSICOCHEMICAL
RP PROPERTIES.
RX PubMed=9644262; DOI=10.1093/oxfordjournals.jbchem.a022078;
RA Mine T., Morita Y., Kataoka A., Mizushima T., Tsuchiya T.;
RT "Evidence for chloramphenicol/H+ antiport in Cmr (MdfA) system of
RT Escherichia coli and properties of the antiporter.";
RL J. Biochem. 124:187-193(1998).
RN [8]
RP TOPOLOGY.
RX PubMed=10022825; DOI=10.1093/emboj/18.4.822;
RA Edgar R., Bibi E.;
RT "A single membrane-embedded negative charge is critical for recognizing
RT positively charged drugs by the Escherichia coli multidrug resistance
RT protein MdfA.";
RL EMBO J. 18:822-832(1999).
RN [9]
RP FUNCTION.
RX PubMed=12578981; DOI=10.1073/pnas.0435544100;
RA Lewinson O., Adler J., Poelarends G.J., Mazurkiewicz P., Driessen A.J.,
RA Bibi E.;
RT "The Escherichia coli multidrug transporter MdfA catalyzes both
RT electrogenic and electroneutral transport reactions.";
RL Proc. Natl. Acad. Sci. U.S.A. 100:1667-1672(2003).
RN [10]
RP MUTAGENESIS OF GLU-26.
RX PubMed=14717607; DOI=10.1021/bi035485t;
RA Adler J., Lewinson O., Bibi E.;
RT "Role of a conserved membrane-embedded acidic residue in the multidrug
RT transporter MdfA.";
RL Biochemistry 43:518-525(2004).
RN [11]
RP FUNCTION IN ALKALITOLERANCE.
RX PubMed=15371593; DOI=10.1073/pnas.0405375101;
RA Lewinson O., Padan E., Bibi E.;
RT "Alkalitolerance: a biological function for a multidrug transporter in pH
RT homeostasis.";
RL Proc. Natl. Acad. Sci. U.S.A. 101:14073-14078(2004).
RN [12]
RP TOPOLOGY [LARGE SCALE ANALYSIS].
RC STRAIN=K12 / MG1655 / ATCC 47076;
RX PubMed=15919996; DOI=10.1126/science.1109730;
RA Daley D.O., Rapp M., Granseth E., Melen K., Drew D., von Heijne G.;
RT "Global topology analysis of the Escherichia coli inner membrane
RT proteome.";
RL Science 308:1321-1323(2005).
RN [13]
RP SUBUNIT.
RX PubMed=17407265; DOI=10.1021/bi602405w;
RA Sigal N., Lewinson O., Wolf S.G., Bibi E.;
RT "E. coli multidrug transporter MdfA is a monomer.";
RL Biochemistry 46:5200-5208(2007).
RN [14]
RP SUBSTRATE-BINDING.
RX PubMed=19808670; DOI=10.1074/jbc.m109.050658;
RA Fluman N., Cohen-Karni D., Weiss T., Bibi E.;
RT "A promiscuous conformational switch in the secondary multidrug transporter
RT MdfA.";
RL J. Biol. Chem. 284:32296-32304(2009).
CC -!- FUNCTION: Efflux pump driven by the proton motive force. Confers
CC resistance to a broad spectrum of chemically unrelated drugs. Confers
CC resistance to a diverse group of cationic or zwitterionic lipophilic
CC compounds such as ethidium bromide, tetraphenylphosphonium, rhodamine,
CC daunomycin, benzalkonium, rifampicin, tetracycline, puromycin, and to
CC chemically unrelated, clinically important antibiotics such as
CC chloramphenicol, erythromycin, and certain aminoglycosides and
CC fluoroquinolones. Overexpression results in isopropyl-beta-D-
CC thiogalactopyranoside (IPTG) exclusion and spectinomycin sensitivity.
CC Transport of neutral substrates is electrogenic, whereas transport of
CC cationic substrates is electroneutral. In addition to its role in
CC multidrug resistance, confers extreme alkaline pH resistance, allowing
CC the growth under conditions that are close to those used normally by
CC alkaliphiles. This activity requires Na(+) or K(+).
CC {ECO:0000269|PubMed:12578981, ECO:0000269|PubMed:15371593,
CC ECO:0000269|PubMed:9079913, ECO:0000269|PubMed:9644262,
CC ECO:0000269|PubMed:9811673}.
CC -!- ACTIVITY REGULATION: Inhibited by CCCP. {ECO:0000269|PubMed:9644262}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC pH dependence:
CC Optimum pH is 6.5. {ECO:0000269|PubMed:9644262};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:17407265}.
CC -!- SUBCELLULAR LOCATION: Cell inner membrane; Multi-pass membrane protein.
CC -!- MISCELLANEOUS: A negative charge at position 26 is required for
CC efficient transport of positively charged substrates, but not for
CC neutral compounds. This negative charge does not play an essential role
CC in the multidrug transport mechanism.
CC -!- MISCELLANEOUS: Has a sensitive conformational switch that can be
CC triggered either by substrate binding or by attaching unrelated agents
CC inside the pocket. Dissimilar substrates induce similar conformational
CC changes.
CC -!- SIMILARITY: Belongs to the major facilitator superfamily. MdfA family.
CC {ECO:0000305}.
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DR EMBL; U44900; AAC44147.1; -; Genomic_DNA.
DR EMBL; Y08743; CAA69997.1; -; Genomic_DNA.
DR EMBL; U00096; AAC73929.1; -; Genomic_DNA.
DR EMBL; AP009048; BAA35546.1; -; Genomic_DNA.
DR PIR; B64822; B64822.
DR RefSeq; NP_415363.1; NC_000913.3.
DR RefSeq; WP_001180089.1; NZ_SSZK01000002.1.
DR PDB; 4ZOW; X-ray; 2.45 A; A=10-400.
DR PDB; 4ZP0; X-ray; 2.00 A; A=9-400.
DR PDB; 4ZP2; X-ray; 2.20 A; A=9-400.
DR PDB; 6EUQ; X-ray; 2.20 A; A=1-410.
DR PDB; 6GV1; X-ray; 3.40 A; A=1-410.
DR PDB; 6OOM; X-ray; 2.20 A; A=9-400.
DR PDB; 6OOP; X-ray; 2.80 A; A=9-400.
DR PDB; 6OOQ; X-ray; 3.00 A; A=9-400.
DR PDB; 6VRZ; X-ray; 2.00 A; A=14-400.
DR PDB; 6VS0; X-ray; 2.10 A; A=14-400.
DR PDB; 6VS1; X-ray; 3.00 A; A=14-400.
DR PDB; 6VS2; X-ray; 3.00 A; A=14-400.
DR PDBsum; 4ZOW; -.
DR PDBsum; 4ZP0; -.
DR PDBsum; 4ZP2; -.
DR PDBsum; 6EUQ; -.
DR PDBsum; 6GV1; -.
DR PDBsum; 6OOM; -.
DR PDBsum; 6OOP; -.
DR PDBsum; 6OOQ; -.
DR PDBsum; 6VRZ; -.
DR PDBsum; 6VS0; -.
DR PDBsum; 6VS1; -.
DR PDBsum; 6VS2; -.
DR AlphaFoldDB; P0AEY8; -.
DR SMR; P0AEY8; -.
DR BioGRID; 4262828; 108.
DR DIP; DIP-48116N; -.
DR IntAct; P0AEY8; 1.
DR STRING; 511145.b0842; -.
DR BindingDB; P0AEY8; -.
DR ChEMBL; CHEMBL4785; -.
DR TCDB; 2.A.1.2.19; the major facilitator superfamily (mfs).
DR PaxDb; P0AEY8; -.
DR PRIDE; P0AEY8; -.
DR ABCD; P0AEY8; 1 sequenced antibody.
DR EnsemblBacteria; AAC73929; AAC73929; b0842.
DR EnsemblBacteria; BAA35546; BAA35546; BAA35546.
DR GeneID; 945448; -.
DR KEGG; ag:CAA69997; -.
DR KEGG; ecj:JW0826; -.
DR KEGG; eco:b0842; -.
DR PATRIC; fig|1411691.4.peg.1436; -.
DR EchoBASE; EB3460; -.
DR eggNOG; COG2814; Bacteria.
DR HOGENOM; CLU_001265_47_2_6; -.
DR InParanoid; P0AEY8; -.
DR OMA; AVLYRMT; -.
DR PhylomeDB; P0AEY8; -.
DR BioCyc; EcoCyc:CMR-MON; -.
DR BioCyc; MetaCyc:CMR-MON; -.
DR BRENDA; 7.6.2.2; 2026.
DR PRO; PR:P0AEY8; -.
DR Proteomes; UP000000318; Chromosome.
DR Proteomes; UP000000625; Chromosome.
DR GO; GO:0005887; C:integral component of plasma membrane; IDA:EcoCyc.
DR GO; GO:0005886; C:plasma membrane; IDA:EcoCyc.
DR GO; GO:0015386; F:potassium:proton antiporter activity; IDA:EcoCyc.
DR GO; GO:0015385; F:sodium:proton antiporter activity; IDA:EcoCyc.
DR GO; GO:0022857; F:transmembrane transporter activity; IBA:GO_Central.
DR GO; GO:0030641; P:regulation of cellular pH; IMP:EcoCyc.
DR GO; GO:0046677; P:response to antibiotic; IMP:EcoCyc.
DR GO; GO:0055085; P:transmembrane transport; IBA:GO_Central.
DR GO; GO:1990961; P:xenobiotic detoxification by transmembrane export across the plasma membrane; IDA:EcoCyc.
DR InterPro; IPR011701; MFS.
DR InterPro; IPR020846; MFS_dom.
DR InterPro; IPR036259; MFS_trans_sf.
DR InterPro; IPR005829; Sugar_transporter_CS.
DR Pfam; PF07690; MFS_1; 1.
DR SUPFAM; SSF103473; SSF103473; 1.
DR PROSITE; PS50850; MFS; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; Cell inner membrane; Cell membrane;
KW Membrane; Reference proteome; Transmembrane; Transmembrane helix;
KW Transport.
FT CHAIN 1..410
FT /note="Multidrug transporter MdfA"
FT /id="PRO_0000173331"
FT TOPO_DOM 1..14
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 15..32
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 33..53
FT /note="Periplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 54..72
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 73..82
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 83..103
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 104..109
FT /note="Periplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 110..130
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 131..144
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 145..165
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 166
FT /note="Periplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 167..187
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 188..226
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 227..247
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 248..255
FT /note="Periplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 256..276
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 277..287
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 288..308
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 309..314
FT /note="Periplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 315..335
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 336..346
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 347..367
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 368..378
FT /note="Periplasmic"
FT /evidence="ECO:0000305"
FT TRANSMEM 379..399
FT /note="Helical"
FT /evidence="ECO:0000305"
FT TOPO_DOM 400..410
FT /note="Cytoplasmic"
FT /evidence="ECO:0000305"
FT MUTAGEN 26
FT /note="E->A: Loss of ethidium bromide efflux activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->D: No change in ethidium bromide efflux
FT activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->H: Almost no chloramphenicol efflux activity.
FT Loss of ethidium bromide efflux activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->I,V: Slight decrease in chloramphenicol efflux
FT activity. Exhibits low ethidium bromide efflux activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->K: Decrease in TPP efflux activity. Exhibits low
FT ethidium bromide efflux activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->L,N: Strong decrease in chloramphenicol efflux
FT activity. Loss of ethidium bromide and TPP efflux
FT activities."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->Q: Slight decrease in chloramphenicol efflux
FT activity. Loss of ethidium bromide efflux activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT MUTAGEN 26
FT /note="E->T: Strong decrease in chloramphenicol efflux
FT activity. Loss of ethidium bromide efflux activity."
FT /evidence="ECO:0000269|PubMed:14717607"
FT CONFLICT 59
FT /note="T -> N (in Ref. 2; CAA69997)"
FT /evidence="ECO:0000305"
FT CONFLICT 225..240
FT /note="ALALGFVSLPLLAWIA -> GAGAADSLVCRCWRGSP (in Ref. 1;
FT AAC44147)"
FT /evidence="ECO:0000305"
FT STRAND 11..13
FT /evidence="ECO:0007829|PDB:4ZP2"
FT HELIX 14..16
FT /evidence="ECO:0007829|PDB:4ZP2"
FT HELIX 17..34
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 36..46
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 53..65
FT /evidence="ECO:0007829|PDB:4ZP0"
FT TURN 66..68
FT /evidence="ECO:0007829|PDB:6GV1"
FT HELIX 69..79
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 81..98
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 99..101
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 105..116
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 117..120
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 121..124
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 126..133
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 136..165
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 170..191
FT /evidence="ECO:0007829|PDB:4ZP0"
FT TURN 197..200
FT /evidence="ECO:0007829|PDB:6OOP"
FT HELIX 205..216
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 219..246
FT /evidence="ECO:0007829|PDB:4ZP0"
FT TURN 247..249
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 254..278
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 279..281
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 284..308
FT /evidence="ECO:0007829|PDB:4ZP0"
FT HELIX 313..339
FT /evidence="ECO:0007829|PDB:4ZP0"
FT STRAND 342..344
FT /evidence="ECO:0007829|PDB:6OOQ"
FT HELIX 346..398
FT /evidence="ECO:0007829|PDB:4ZP0"
SQ SEQUENCE 410 AA; 44321 MW; D4466EE680095773 CRC64;
MQNKLASGAR LGRQALLFPL CLVLYEFSTY IGNDMIQPGM LAVVEQYQAG IDWVPTSMTA
YLAGGMFLQW LLGPLSDRIG RRPVMLAGVV WFIVTCLAIL LAQNIEQFTL LRFLQGISLC
FIGAVGYAAI QESFEEAVCI KITALMANVA LIAPLLGPLV GAAWIHVLPW EGMFVLFAAL
AAISFFGLQR AMPETATRIG EKLSLKELGR DYKLVLKNGR FVAGALALGF VSLPLLAWIA
QSPIIIITGE QLSSYEYGLL QVPIFGALIA GNLLLARLTS RRTVRSLIIM GGWPIMIGLL
VAAAATVISS HAYLWMTAGL SIYAFGIGLA NAGLVRLTLF ASDMSKGTVS AAMGMLQMLI
FTVGIEISKH AWLNGGNGLF NLFNLVNGIL WLSLMVIFLK DKQMGNSHEG
