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Protein

Multidrug transporter MdfA

Gene

mdfA

Organism
Escherichia coli (strain K12)
Status
Reviewed-Annotation score: Annotation score: 5 out of 5-Experimental evidence at protein leveli

Functioni

Efflux pump driven by the proton motive force. Confers resistance to a broad spectrum of chemically unrelated drugs. Confers resistance to a diverse group of cationic or zwitterionic lipophilic compounds such as ethidium bromide, tetraphenylphosphonium, rhodamine, daunomycin, benzalkonium, rifampicin, tetracycline, puromycin, and to chemically unrelated, clinically important antibiotics such as chloramphenicol, erythromycin, and certain aminoglycosides and fluoroquinolones. Overexpression results in isopropyl-beta-D-thiogalactopyranoside (IPTG) exclusion and spectinomycin sensitivity. Transport of neutral substrates is electrogenic, whereas transport of cationic substrates is electroneutral. In addition to its role in multidrug resistance, confers extreme alkaline pH resistance, allowing the growth under conditions that are close to those used normally by alkaliphiles. This activity requires Na+ or K+.5 Publications

Enzyme regulationi

Inhibited by CCCP.1 Publication

pH dependencei

Optimum pH is 6.5.1 Publication

GO - Molecular functioni

  • drug transmembrane transporter activity Source: EcoCyc
  • potassium:proton antiporter activity Source: EcoCyc
  • sodium:proton antiporter activity Source: EcoCyc

GO - Biological processi

  • drug transmembrane transport Source: EcoCyc
  • potassium ion transmembrane transport Source: GOC
  • regulation of cellular pH Source: EcoCyc
  • response to antibiotic Source: EcoCyc
  • sodium ion transmembrane transport Source: GOC
Complete GO annotation...

Keywords - Biological processi

Antibiotic resistance, Transport

Enzyme and pathway databases

BioCyciEcoCyc:CMR-MONOMER.
ECOL316407:JW0826-MONOMER.
MetaCyc:CMR-MONOMER.

Protein family/group databases

TCDBi2.A.1.2.19. the major facilitator superfamily (mfs).

Names & Taxonomyi

Protein namesi
Recommended name:
Multidrug transporter MdfA
Alternative name(s):
Chloramphenicol resistance pump Cmr
Gene namesi
Name:mdfA
Synonyms:cmlA, cmr
Ordered Locus Names:b0842, JW0826
OrganismiEscherichia coli (strain K12)
Taxonomic identifieri83333 [NCBI]
Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
Proteomesi
  • UP000000318 Componenti: Chromosome
  • UP000000625 Componenti: Chromosome

Organism-specific databases

EcoGeneiEG13696. mdfA.

Subcellular locationi

Topology

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Topological domaini1 – 1414CytoplasmicCuratedAdd
BLAST
Transmembranei15 – 3218HelicalCuratedAdd
BLAST
Topological domaini33 – 5321PeriplasmicCuratedAdd
BLAST
Transmembranei54 – 7219HelicalCuratedAdd
BLAST
Topological domaini73 – 8210CytoplasmicCurated
Transmembranei83 – 10321HelicalCuratedAdd
BLAST
Topological domaini104 – 1096PeriplasmicCurated
Transmembranei110 – 13021HelicalCuratedAdd
BLAST
Topological domaini131 – 14414CytoplasmicCuratedAdd
BLAST
Transmembranei145 – 16521HelicalCuratedAdd
BLAST
Topological domaini166 – 1661PeriplasmicCurated
Transmembranei167 – 18721HelicalCuratedAdd
BLAST
Topological domaini188 – 22639CytoplasmicCuratedAdd
BLAST
Transmembranei227 – 24721HelicalCuratedAdd
BLAST
Topological domaini248 – 2558PeriplasmicCurated
Transmembranei256 – 27621HelicalCuratedAdd
BLAST
Topological domaini277 – 28711CytoplasmicCuratedAdd
BLAST
Transmembranei288 – 30821HelicalCuratedAdd
BLAST
Topological domaini309 – 3146PeriplasmicCurated
Transmembranei315 – 33521HelicalCuratedAdd
BLAST
Topological domaini336 – 34611CytoplasmicCuratedAdd
BLAST
Transmembranei347 – 36721HelicalCuratedAdd
BLAST
Topological domaini368 – 37811PeriplasmicCuratedAdd
BLAST
Transmembranei379 – 39921HelicalCuratedAdd
BLAST
Topological domaini400 – 41011CytoplasmicCuratedAdd
BLAST

GO - Cellular componenti

  • integral component of plasma membrane Source: EcoCyc
  • plasma membrane Source: EcoCyc
Complete GO annotation...

Keywords - Cellular componenti

Cell inner membrane, Cell membrane, Membrane

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi26 – 261E → A: Loss of ethidium bromide efflux activity. 1 Publication
Mutagenesisi26 – 261E → D: No change in ethidium bromide efflux activity. 1 Publication
Mutagenesisi26 – 261E → H: Almost no chloramphenicol efflux activity. Loss of ethidium bromide efflux activity. 1 Publication
Mutagenesisi26 – 261E → I or V: Slight decrease in chloramphenicol efflux activity. Exhibits low ethidium bromide efflux activity. 1 Publication
Mutagenesisi26 – 261E → K: Decrease in TPP efflux activity. Exhibits low ethidium bromide efflux activity. 1 Publication
Mutagenesisi26 – 261E → L or N: Strong decrease in chloramphenicol efflux activity. Loss of ethidium bromide and TPP efflux activities. 1 Publication
Mutagenesisi26 – 261E → Q: Slight decrease in chloramphenicol efflux activity. Loss of ethidium bromide efflux activity. 1 Publication
Mutagenesisi26 – 261E → T: Strong decrease in chloramphenicol efflux activity. Loss of ethidium bromide efflux activity. 1 Publication

Chemistry

ChEMBLiCHEMBL4785.

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 410410Multidrug transporter MdfAPRO_0000173331Add
BLAST

Proteomic databases

PaxDbiP0AEY8.

Interactioni

Subunit structurei

Monomer.1 Publication

Protein-protein interaction databases

BioGridi4262828. 106 interactions.
DIPiDIP-48116N.
IntActiP0AEY8. 1 interaction.
MINTiMINT-1227586.
STRINGi511145.b0842.

Chemistry

BindingDBiP0AEY8.

Structurei

Secondary structure

1
410
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi11 – 133Combined sources
Helixi14 – 163Combined sources
Helixi17 – 3418Combined sources
Helixi36 – 4611Combined sources
Helixi53 – 6513Combined sources
Helixi69 – 7911Combined sources
Helixi81 – 9818Combined sources
Helixi99 – 1013Combined sources
Helixi105 – 11612Combined sources
Helixi117 – 1204Combined sources
Helixi121 – 1244Combined sources
Helixi126 – 1338Combined sources
Helixi136 – 16530Combined sources
Helixi170 – 19122Combined sources
Helixi205 – 21612Combined sources
Helixi219 – 24628Combined sources
Turni247 – 2493Combined sources
Helixi254 – 27825Combined sources
Helixi279 – 2813Combined sources
Helixi284 – 30825Combined sources
Helixi313 – 33927Combined sources
Helixi346 – 39853Combined sources

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
4ZOWX-ray2.45A10-400[»]
4ZP0X-ray2.00A9-400[»]
4ZP2X-ray2.20A9-400[»]
ProteinModelPortaliP0AEY8.
SMRiP0AEY8. Positions 9-400.
ModBaseiSearch...
MobiDBiSearch...

Family & Domainsi

Sequence similaritiesi

Keywords - Domaini

Transmembrane, Transmembrane helix

Phylogenomic databases

eggNOGiENOG4107QF7. Bacteria.
ENOG410XT98. LUCA.
HOGENOMiHOG000118215.
InParanoidiP0AEY8.
KOiK08160.
OMAiMQTHANR.
OrthoDBiEOG6NPM4Z.
PhylomeDBiP0AEY8.

Family and domain databases

InterProiIPR011701. MFS.
IPR020846. MFS_dom.
IPR005829. Sugar_transporter_CS.
[Graphical view]
PfamiPF07690. MFS_1. 1 hit.
[Graphical view]
SUPFAMiSSF103473. SSF103473. 1 hit.
PROSITEiPS50850. MFS. 1 hit.
[Graphical view]

Sequencei

Sequence statusi: Complete.

P0AEY8-1 [UniParc]FASTAAdd to basket

« Hide

        10         20         30         40         50
MQNKLASGAR LGRQALLFPL CLVLYEFSTY IGNDMIQPGM LAVVEQYQAG
60 70 80 90 100
IDWVPTSMTA YLAGGMFLQW LLGPLSDRIG RRPVMLAGVV WFIVTCLAIL
110 120 130 140 150
LAQNIEQFTL LRFLQGISLC FIGAVGYAAI QESFEEAVCI KITALMANVA
160 170 180 190 200
LIAPLLGPLV GAAWIHVLPW EGMFVLFAAL AAISFFGLQR AMPETATRIG
210 220 230 240 250
EKLSLKELGR DYKLVLKNGR FVAGALALGF VSLPLLAWIA QSPIIIITGE
260 270 280 290 300
QLSSYEYGLL QVPIFGALIA GNLLLARLTS RRTVRSLIIM GGWPIMIGLL
310 320 330 340 350
VAAAATVISS HAYLWMTAGL SIYAFGIGLA NAGLVRLTLF ASDMSKGTVS
360 370 380 390 400
AAMGMLQMLI FTVGIEISKH AWLNGGNGLF NLFNLVNGIL WLSLMVIFLK
410
DKQMGNSHEG
Length:410
Mass (Da):44,321
Last modified:December 20, 2005 - v1
Checksum:iD4466EE680095773
GO

Experimental Info

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Sequence conflicti59 – 591T → N in CAA69997 (PubMed:9079913).Curated
Sequence conflicti225 – 24016ALALG…LAWIA → GAGAADSLVCRCWRGSP in AAC44147 (PubMed:8655497).CuratedAdd
BLAST

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
U44900 Genomic DNA. Translation: AAC44147.1.
Y08743 Genomic DNA. Translation: CAA69997.1.
U00096 Genomic DNA. Translation: AAC73929.1.
AP009048 Genomic DNA. Translation: BAA35546.1.
PIRiB64822.
RefSeqiNP_415363.1. NC_000913.3.
WP_001180089.1. NZ_LN832404.1.

Genome annotation databases

EnsemblBacteriaiAAC73929; AAC73929; b0842.
BAA35546; BAA35546; BAA35546.
GeneIDi945448.
KEGGiag:CAA69997.
ecj:JW0826.
eco:b0842.
PATRICi32116889. VBIEscCol129921_0870.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBLi
GenBanki
DDBJi
Links Updated
U44900 Genomic DNA. Translation: AAC44147.1.
Y08743 Genomic DNA. Translation: CAA69997.1.
U00096 Genomic DNA. Translation: AAC73929.1.
AP009048 Genomic DNA. Translation: BAA35546.1.
PIRiB64822.
RefSeqiNP_415363.1. NC_000913.3.
WP_001180089.1. NZ_LN832404.1.

3D structure databases

Select the link destinations:
PDBei
RCSB PDBi
PDBji
Links Updated
EntryMethodResolution (Å)ChainPositionsPDBsum
4ZOWX-ray2.45A10-400[»]
4ZP0X-ray2.00A9-400[»]
4ZP2X-ray2.20A9-400[»]
ProteinModelPortaliP0AEY8.
SMRiP0AEY8. Positions 9-400.
ModBaseiSearch...
MobiDBiSearch...

Protein-protein interaction databases

BioGridi4262828. 106 interactions.
DIPiDIP-48116N.
IntActiP0AEY8. 1 interaction.
MINTiMINT-1227586.
STRINGi511145.b0842.

Chemistry

BindingDBiP0AEY8.
ChEMBLiCHEMBL4785.

Protein family/group databases

TCDBi2.A.1.2.19. the major facilitator superfamily (mfs).

Proteomic databases

PaxDbiP0AEY8.

Protocols and materials databases

Structural Biology KnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaiAAC73929; AAC73929; b0842.
BAA35546; BAA35546; BAA35546.
GeneIDi945448.
KEGGiag:CAA69997.
ecj:JW0826.
eco:b0842.
PATRICi32116889. VBIEscCol129921_0870.

Organism-specific databases

EchoBASEiEB3460.
EcoGeneiEG13696. mdfA.

Phylogenomic databases

eggNOGiENOG4107QF7. Bacteria.
ENOG410XT98. LUCA.
HOGENOMiHOG000118215.
InParanoidiP0AEY8.
KOiK08160.
OMAiMQTHANR.
OrthoDBiEOG6NPM4Z.
PhylomeDBiP0AEY8.

Enzyme and pathway databases

BioCyciEcoCyc:CMR-MONOMER.
ECOL316407:JW0826-MONOMER.
MetaCyc:CMR-MONOMER.

Miscellaneous databases

PROiP0AEY8.

Family and domain databases

InterProiIPR011701. MFS.
IPR020846. MFS_dom.
IPR005829. Sugar_transporter_CS.
[Graphical view]
PfamiPF07690. MFS_1. 1 hit.
[Graphical view]
SUPFAMiSSF103473. SSF103473. 1 hit.
PROSITEiPS50850. MFS. 1 hit.
[Graphical view]
ProtoNetiSearch...

Publicationsi

« Hide 'large scale' publications
  1. "Isolation of cmr, a novel Escherichia coli chloramphenicol resistance gene encoding a putative efflux pump."
    Nilsen I.W., Bakke I., Vader A., Olsvik O., El-Gewely M.R.
    J. Bacteriol. 178:3188-3193(1996) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
    Strain: K12.
  2. "MdfA, an Escherichia coli multidrug resistance protein with an extraordinarily broad spectrum of drug recognition."
    Edgar R., Bibi E.
    J. Bacteriol. 179:2274-2280(1997) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION.
    Strain: ATCC 33694 / HB101.
  3. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
    Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
  4. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
    Strain: K12 / MG1655 / ATCC 47076.
  5. "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655 and W3110."
    Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S., Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.
    Mol. Syst. Biol. 2:E1-E5(2006) [PubMed] [Europe PMC] [Abstract]
    Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
    Strain: K12 / W3110 / ATCC 27325 / DSM 5911.
  6. "The Escherichia coli cmlA gene encodes the multidrug efflux pump Cmr/MdfA and is responsible for isopropyl-beta-D-thiogalactopyranoside exclusion and spectinomycin sensitivity."
    Bohn C., Bouloc P.
    J. Bacteriol. 180:6072-6075(1998) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION.
    Strain: K12 / C600 / ATCC 23724 / DSM 3925 / LMG 3041 / NCIB 10222.
  7. "Evidence for chloramphenicol/H+ antiport in Cmr (MdfA) system of Escherichia coli and properties of the antiporter."
    Mine T., Morita Y., Kataoka A., Mizushima T., Tsuchiya T.
    J. Biochem. 124:187-193(1998) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION AS AN ANTIPORTER, ENZYME REGULATION, BIOPHYSICOCHEMICAL PROPERTIES.
  8. "A single membrane-embedded negative charge is critical for recognizing positively charged drugs by the Escherichia coli multidrug resistance protein MdfA."
    Edgar R., Bibi E.
    EMBO J. 18:822-832(1999) [PubMed] [Europe PMC] [Abstract]
    Cited for: TOPOLOGY.
  9. "The Escherichia coli multidrug transporter MdfA catalyzes both electrogenic and electroneutral transport reactions."
    Lewinson O., Adler J., Poelarends G.J., Mazurkiewicz P., Driessen A.J., Bibi E.
    Proc. Natl. Acad. Sci. U.S.A. 100:1667-1672(2003) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION.
  10. "Role of a conserved membrane-embedded acidic residue in the multidrug transporter MdfA."
    Adler J., Lewinson O., Bibi E.
    Biochemistry 43:518-525(2004) [PubMed] [Europe PMC] [Abstract]
    Cited for: MUTAGENESIS OF GLU-26.
  11. "Alkalitolerance: a biological function for a multidrug transporter in pH homeostasis."
    Lewinson O., Padan E., Bibi E.
    Proc. Natl. Acad. Sci. U.S.A. 101:14073-14078(2004) [PubMed] [Europe PMC] [Abstract]
    Cited for: FUNCTION IN ALKALITOLERANCE.
  12. "Global topology analysis of the Escherichia coli inner membrane proteome."
    Daley D.O., Rapp M., Granseth E., Melen K., Drew D., von Heijne G.
    Science 308:1321-1323(2005) [PubMed] [Europe PMC] [Abstract]
    Cited for: TOPOLOGY [LARGE SCALE ANALYSIS].
    Strain: K12 / MG1655 / ATCC 47076.
  13. "E. coli multidrug transporter MdfA is a monomer."
    Sigal N., Lewinson O., Wolf S.G., Bibi E.
    Biochemistry 46:5200-5208(2007) [PubMed] [Europe PMC] [Abstract]
    Cited for: SUBUNIT.
  14. "A promiscuous conformational switch in the secondary multidrug transporter MdfA."
    Fluman N., Cohen-Karni D., Weiss T., Bibi E.
    J. Biol. Chem. 284:32296-32304(2009) [PubMed] [Europe PMC] [Abstract]
    Cited for: SUBSTRATE-BINDING.

Entry informationi

Entry nameiMDFA_ECOLI
AccessioniPrimary (citable) accession number: P0AEY8
Secondary accession number(s): P71226, P75807, Q46966
Entry historyi
Integrated into UniProtKB/Swiss-Prot: December 20, 2005
Last sequence update: December 20, 2005
Last modified: January 20, 2016
This is version 90 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Miscellaneous

A negative charge at position 26 is required for efficient transport of positively charged substrates, but not for neutral compounds. This negative charge does not play an essential role in the multidrug transport mechanism.
Has a sensitive conformational switch that can be triggered either by substrate binding or by attaching unrelated agents inside the pocket. Dissimilar substrates induce similar conformational changes.

Keywords - Technical termi

3D-structure, Complete proteome, Reference proteome

Documents

  1. Escherichia coli
    Escherichia coli (strain K12): entries and cross-references to EcoGene
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references
  3. SIMILARITY comments
    Index of protein domains and families

Similar proteinsi

Links to similar proteins from the UniProt Reference Clusters (UniRef) at 100%, 90% and 50% sequence identity:
100%UniRef100 combines identical sequences and sub-fragments with 11 or more residues from any organism into one UniRef entry.
90%UniRef90 is built by clustering UniRef100 sequences that have at least 90% sequence identity to, and 80% overlap with, the longest sequence (a.k.a seed sequence).
50%UniRef50 is built by clustering UniRef90 seed sequences that have at least 50% sequence identity to, and 80% overlap with, the longest sequence in the cluster.