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Escherichia coli K-12 substr. MG1655 Transporter: Zn2+ / Fe2+ / Cd2+ efflux transporter FieF



Gene: fieF Accession Numbers: EG11873 (EcoCyc), b3915, ECK3907

Synonyms: yiiP

Regulation Summary Diagram: ?

Subunit composition of Zn2+ / Fe2+ / Cd2+ efflux transporter FieF = [FieF]2

Summary:
The FieF protein is a member of the cation diffusion facilitator (CDF) family of metal cation transporters [Paulsen97a, Saier14]. FieF functions as a divalent metal cation transporter which exports Zn2+, Cd2+, and possibly Fe2+ but probably not Hg2+, Co2+, Ni2+, Mn2+, Ca2+ or Mg2+. Metal binding properties of the purified protein identifying zinc and cadmium as substrates have been described [Chao04, Wei05, Wei06].

FieF forms dimers in detergent-lipid micelles and in reconstituted membranes [Wei04]. The structure of FieF in complex with zinc has been determined by X-ray crystallography to a resolution of 3.8 Å. The FieF homodimer forms a 'Y'-shaped molecule in perpendicular orientation to the plane of the membrane. The transmembrane domains (TMD) of each monomer form a six helix compact bundle; the C-terminal domains (CTD) protrude into the cytoplasm and their association provides the stem of the 'Y'. Two cavities are identifed in the structure - an extracellular cavity exposed to the membrane outer leaflet and an intracellular cavity confined between the TMD and CTD at the inner face of the membrane [Lu07a]. The CTD adopts a metallochaperone-like fold and may regulate metal transport activity [Lu09a]. Leucine residue 152 constitutes the principle hydrophobic barrier between the two cavities; a putative transmembrane zinc pathway runs from the intracellular cavity through L152 within the inter-cavity seal to the transport site in the extracellular cavity; protein conformational change alternates the on-off mode of zinc binding and protonation-deprotonation of the transport site in a coordinated manner [Lu09a]

E.coli cells overexpressing fieF exhibited reduced isotopic Fe accumulation, and Zn(II) efflux was shown to be catalyzed through a proton antiport mechanism in membrane eversion studies. Transmembrane flux of iron cations was measured using fluorescence quenching in conjunction with purified and reconstituted FieF. The results suggest that FieF is an iron and zinc efflux system which functions in detoxification [Grass05].

The fieF gene probably constitutes a monocistronic operon. Transcription of fieF has been shown to correlate with iron concentration independent of the ferrous iron uptake regulator Fur. Double fieF and fur led to growth deficiency in complex growth medium, which was partially alleviated by in trans expression of fieF [Grass05]. Experiments with transcriptional fusions showed fieF is strongly induced by zinc and to a lesser extent by cadmium [Grass01].

FieF: ferrous iron efflux

Citations: [Nies07, Cubillas13]

Locations: inner membrane

Map Position: [4,104,492 -> 4,105,394] (88.47 centisomes)
Length: 903 bp / 300 aa

Molecular Weight of Polypeptide: 32.927 kD (from nucleotide sequence), 25.0 kD (experimental) [Wei04 ]

Molecular Weight of Multimer: 68.0 kD (experimental) [Wei04]

Unification Links: ASAP:ABE-0012786 , DIP:DIP-48564N , EchoBASE:EB1819 , EcoGene:EG11873 , EcoliWiki:b3915 , OU-Microarray:b3915 , PortEco:fieF , PR:PRO_000022615 , Protein Model Portal:P69380 , RefSeq:NP_418350 , RegulonDB:EG11873 , SMR:P69380 , String:511145.b3915 , UniProt:P69380

Relationship Links: InterPro:IN-FAMILY:IPR002524 , InterPro:IN-FAMILY:IPR023783 , InterPro:IN-FAMILY:IPR027469 , InterPro:IN-FAMILY:IPR027470 , Panther:IN-FAMILY:PTHR11562 , PDB:Structure:2qfi , PDB:Structure:3H90 , Pfam:IN-FAMILY:PF01545

In Paralogous Gene Group: 561 (2 members)

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0006829 - zinc ion transport Inferred from experiment Inferred by computational analysis [GOA01a, Grass05]
GO:0006876 - cellular cadmium ion homeostasis Inferred from experiment [Wei05]
GO:0006879 - cellular iron ion homeostasis Inferred from experiment [Grass05]
GO:0006882 - cellular zinc ion homeostasis Inferred from experiment [Grass05]
GO:0015684 - ferrous iron transport Inferred from experiment Inferred by computational analysis [GOA01a, Grass05]
GO:0015691 - cadmium ion transport Inferred from experiment [Wei05]
GO:0070574 - cadmium ion transmembrane transport Inferred from experiment [Wei05]
GO:0071577 - zinc ion transmembrane transport Inferred from experiment [Grass05]
GO:0098655 - cation transmembrane transport Inferred by computational analysis Inferred from experiment [Grass05, GOA01a]
GO:0006810 - transport Inferred by computational analysis [UniProtGOA11a]
GO:0006811 - ion transport Inferred by computational analysis [UniProtGOA11a]
GO:0055072 - iron ion homeostasis Inferred by computational analysis [UniProtGOA11a]
Molecular Function: GO:0015086 - cadmium ion transmembrane transporter activity Inferred from experiment [Wei05]
GO:0015093 - ferrous iron transmembrane transporter activity Inferred from experiment [Grass05]
GO:0015341 - zinc efflux active transmembrane transporter activity Inferred from experiment [Grass05]
GO:0008324 - cation transmembrane transporter activity Inferred by computational analysis [GOA01a]
GO:0046873 - metal ion transmembrane transporter activity Inferred by computational analysis [GOA01a]
Cellular Component: GO:0005886 - plasma membrane Inferred from experiment Inferred by computational analysis [UniProtGOA11, UniProtGOA11a, DiazMejia09, Daley05, Chao04]
GO:0005887 - integral component of plasma membrane Inferred from experiment [Lu07a]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11a]
GO:0016021 - integral component of membrane Inferred by computational analysis [UniProtGOA11a, GOA01a]

MultiFun Terms: cell structure membrane
transport Electrochemical potential driven transporters Porters (Uni-, Sym- and Antiporters)

Essentiality data for fieF knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Credits:
Created 31-Oct-2007 by Johnson A , JCVI
Curated 31-Oct-2007 by Johnson A , JCVI
Last-Curated ? 11-Aug-2014 by Mackie A , Macquarie University


Enzymatic reaction of: Cd2+:H+ antiporter (Zn2+ / Fe2+ / Cd2+ efflux transporter FieF)

Synonyms: export of cadmium

Kinetic Parameters:

Substrate
Km (μM)
Citations
Cd2+
266.0
[Wei05]


Enzymatic reaction of: Fe2+:H+ antiporter (Zn2+ / Fe2+ / Cd2+ efflux transporter FieF)

Synonyms: export of iron


Enzymatic reaction of: Zn2+:H+ antiporter (Zn2+ / Fe2+ / Cd2+ efflux transporter FieF)

Synonyms: export of zinc

Kinetic Parameters:

Substrate
Km (μM)
Citations
Zn2+
310.0
[Wei05]


Sequence Features

Feature Class Location Common Name Citations Comment
Transmembrane-Region 10 -> 27 TM1
[Lu07a]
 
Transmembrane-Region 12 -> 34  
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 38 -> 60  
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Metal-Binding-Site 45, 49, 153, 157 Z1 binding site
[Wei05, Wei06, Gupta14]
implicated in metal (Zn2+ and Cd2+ coordination
Transmembrane-Region 45 -> 63 TM2
[Lu07a]
 
Transmembrane-Region 78 -> 99 TM3
[Lu07a]
 
Transmembrane-Region 80 -> 102  
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 117 -> 139  
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 127 -> 141 TM4
[Lu07a]
 
Transmembrane-Region 146 -> 159 TM5
[Lu07a]
 
Amino-Acid-Site 152 proton gate
[Gupta14]
L152 serves as the principal hydrophobic barrier in the transport pathway
Active-Site 153  
[Gupta14]
H153 constitutes the protonation site
Transmembrane-Region 158 -> 175  
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 180 -> 199  
[UniProt10a]
UniProt: Helical;; Non-Experimental Qualifier: potential;
Transmembrane-Region 182 -> 205 TM6
[Lu07a]
 


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b3915 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11873; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

Chao04: Chao Y, Fu D (2004). "Thermodynamic studies of the mechanism of metal binding to the Escherichia coli zinc transporter YiiP." J Biol Chem 279(17);17173-80. PMID: 14960568

Cubillas13: Cubillas C, Vinuesa P, Tabche ML, Garcia-de Los Santos A (2013). "Phylogenomic analysis of Cation Diffusion Facilitator proteins uncovers Ni(2+)/Co(2+) transporters." Metallomics. PMID: 24077251

Daley05: Daley DO, Rapp M, Granseth E, Melen K, Drew D, von Heijne G (2005). "Global topology analysis of the Escherichia coli inner membrane proteome." Science 308(5726);1321-3. PMID: 15919996

DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

Grass01: Grass G, Fan B, Rosen BP, Franke S, Nies DH, Rensing C (2001). "ZitB (YbgR), a member of the cation diffusion facilitator family, is an additional zinc transporter in Escherichia coli." J Bacteriol 2001;183(15);4664-7. PMID: 11443104

Grass05: Grass G, Otto M, Fricke B, Haney CJ, Rensing C, Nies DH, Munkelt D (2005). "FieF (YiiP) from Escherichia coli mediates decreased cellular accumulation of iron and relieves iron stress." Arch Microbiol 183(1);9-18. PMID: 15549269

Gupta14: Gupta S, Chai J, Cheng J, D'Mello R, Chance MR, Fu D (2014). "Visualizing the kinetic power stroke that drives proton-coupled zinc(II) transport." Nature 512(7512);101-4. PMID: 25043033

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Lu07a: Lu M, Fu D (2007). "Structure of the zinc transporter YiiP." Science 317(5845);1746-8. PMID: 17717154

Lu09a: Lu M, Chai J, Fu D (2009). "Structural basis for autoregulation of the zinc transporter YiiP." Nat Struct Mol Biol 16(10);1063-7. PMID: 19749753

Nies07: Nies DH (2007). "Biochemistry. How cells control zinc homeostasis." Science 317(5845);1695-6. PMID: 17885121

Paulsen97a: Paulsen IT, Saier MH (1997). "A novel family of ubiquitous heavy metal ion transport proteins." J Membr Biol 1997;156(2);99-103. PMID: 9075641

Saier14: Saier MH, Reddy VS, Tamang DG, Vastermark A (2014). "The transporter classification database." Nucleic Acids Res 42(1);D251-8. PMID: 24225317

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on the manual assignment of UniProtKB Subcellular Location terms in UniProtKB/Swiss-Prot entries."

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

Wei04: Wei Y, Li H, Fu D (2004). "Oligomeric state of the Escherichia coli metal transporter YiiP." J Biol Chem 279(38);39251-9. PMID: 15258151

Wei05: Wei Y, Fu D (2005). "Selective metal binding to a membrane-embedded aspartate in the Escherichia coli metal transporter YiiP (FieF)." J Biol Chem 280(40);33716-24. PMID: 16049012

Wei06: Wei Y, Fu D (2006). "Binding and transport of metal ions at the dimer interface of the Escherichia coli metal transporter YiiP." J Biol Chem 281(33);23492-502. PMID: 16790427


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Sat Dec 20, 2014, biocyc14.