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Escherichia coli K-12 substr. MG1655 Enzyme: regulator of KefC-mediated potassium transport and quinone oxidoreductase



Gene: kefF Accession Numbers: EG11568 (EcoCyc), b0046, ECK0047

Synonyms: yabF

Regulation Summary Diagram: ?

Subunit composition of regulator of KefC-mediated potassium transport and quinone oxidoreductase = [KefF]2
         regulator of KefC-mediated potassium transport and quinone oxidoreductase = KefF

Summary:
KefF is an activator of potassium transport mediated by the KefC antiporter [Miller00a, Fujisawa07]. KefF also has enzymatic activity as a quinone oxidoreductase, thereby apparently reducing the redox toxicity of electrophilic quinones. This enzymatic activity of KefF is not required for its function as an activator of KefC [Lyngberg11]. NAD(P)H dehydrogenase activity of KefF was predicted based on sequence similarity [Reed03].

Crystal structures of an artificially constructed KefF fusion with the C-terminal domain of KefC have been solved [Roosild09, Roosild10]. KefF may exert its effect on the activity of KefC by influencing the angle of KefC's hinge domain [Roosild09].

A kefF mutant exhibits a defect in KefC-mediated potassium transport [Miller00a].

Expression in E. coli of a yabF gene of environmental origin results in carbonyl biosynthesis from polyols [Knietsch03].

Citations: [Miller97]

Locations: cytosol, inner membrane

Map Position: [47,246 -> 47,776] (1.02 centisomes)
Length: 531 bp / 176 aa

Molecular Weight of Polypeptide: 20.17 kD (from nucleotide sequence)

Unification Links: ASAP:ABE-0000159 , DIP:DIP-35822N , EchoBASE:EB1528 , EcoGene:EG11568 , EcoliWiki:b0046 , ModBase:P0A754 , OU-Microarray:b0046 , PortEco:kefF , PR:PRO_000023063 , Protein Model Portal:P0A754 , RefSeq:NP_414588 , RegulonDB:EG11568 , SMR:P0A754 , String:511145.b0046 , UniProt:P0A754

Relationship Links: InterPro:IN-FAMILY:IPR003680 , InterPro:IN-FAMILY:IPR023948 , PDB:Structure:3EYW , PDB:Structure:3L9W , PDB:Structure:3L9X , Pfam:IN-FAMILY:PF02525

In Paralogous Gene Group: 18 (3 members)

Gene-Reaction Schematic: ?

Instance reaction of [a quinone + NAD(P)H + H+ → a quinol + NAD(P)+] (1.6.5.2):
i1: menadione + NADH + H+ = menadiol + NAD+ (1.6.99.5)

GO Terms:

Biological Process: GO:0032414 - positive regulation of ion transmembrane transporter activity Inferred from experiment Inferred by computational analysis [GOA06, Miller00a, Lyngberg11]
GO:0006813 - potassium ion transport Inferred by computational analysis [GOA01a]
GO:0055114 - oxidation-reduction process Inferred by computational analysis [UniProtGOA11a]
GO:1901381 - positive regulation of potassium ion transmembrane transport Inferred by computational analysis [GOA06]
Molecular Function: GO:0003955 - NAD(P)H dehydrogenase (quinone) activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, Lyngberg11]
GO:0009055 - electron carrier activity Inferred from experiment [Fujisawa07]
GO:0010181 - FMN binding Inferred from experiment Inferred by computational analysis [GOA06, Lyngberg11]
GO:0042803 - protein homodimerization activity Inferred from experiment [Lyngberg11]
GO:0016491 - oxidoreductase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]
GO:0005886 - plasma membrane Inferred by computational analysis [UniProtGOA11, UniProtGOA11a]
GO:0016020 - membrane Inferred by computational analysis [UniProtGOA11a]

MultiFun Terms: metabolism
regulation type of regulation posttranscriptional inhibition / activation of enzymes
transport

Essentiality data for kefF 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]

Credits:
Created 11-Aug-2011 by Keseler I , SRI International
Last-Curated ? 11-Aug-2011 by Keseler I , SRI International


Enzymatic reaction of: quinone oxidoreductase

EC Number: 1.6.5.2

a quinone + NAD(P)H + H+ <=> a quinol + NAD(P)+

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is physiologically favored in the direction shown.

Summary:
Benzoquinone, menadione, and duroquinone as well as the 1-electron acceptors ferricyanide and DCIP can all serve as substrates. The enzyme utilizes both NADH and NADPH [Lyngberg11].

Cofactors or Prosthetic Groups: FMN [Lyngberg11]


Sequence Features

Feature Class Location Common Name Attached Group Citations Comment
Amino-Acid-Sites-That-Bind 8    
[UniProt13]
UniProt: FMN.
Nucleotide-Phosphate-Binding-Region 14 -> 17   FMN
[UniProt14]
UniProt: FMN.
Mutagenesis-Variant 41    
[Roosild09, UniProt13]
Alternate sequence: D → K; UniProt: Strongly reduced potassium efflux.
Nucleotide-Phosphate-Binding-Region 65 -> 68   FMN
[UniProt14]
UniProt: FMN.
Nucleotide-Phosphate-Binding-Region 105 -> 108   FMN
[UniProt14]
UniProt: FMN.
Mutagenesis-Variant 107    
[Lyngberg11, UniProt13]
Alternate sequence: G → S; UniProt: Does not bind FMN. Lacks oxidoreductase activity, but is still able to activate potassium efflux.
Mutagenesis-Variant 107 G107S mutant  
[Lyngberg11]
The G107S mutant does not bind FMN and has reduced enzymatic activity.
Mutagenesis-Variant 112    
[Lyngberg11, UniProt13]
Alternate sequence: H → W; UniProt: Lacks oxidoreductase activity, but is still able to activate potassium efflux.
Mutagenesis-Variant 149    
[Lyngberg11, UniProt13]
Alternate sequence: F → W; UniProt: Lacks oxidoreductase activity, but is still able to activate potassium efflux.


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b0046 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11568; 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

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

Fujisawa07: Fujisawa M, Ito M, Krulwich TA (2007). "Three two-component transporters with channel-like properties have monovalent cation/proton antiport activity." Proc Natl Acad Sci U S A 104(33);13289-94. PMID: 17679694

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

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

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

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

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

Knietsch03: Knietsch A, Waschkowitz T, Bowien S, Henne A, Daniel R (2003). "Construction and screening of metagenomic libraries derived from enrichment cultures: generation of a gene bank for genes conferring alcohol oxidoreductase activity on Escherichia coli." Appl Environ Microbiol 69(3);1408-16. PMID: 12620823

Lyngberg11: Lyngberg L, Healy J, Bartlett W, Miller S, Conway SJ, Booth IR, Rasmussen T (2011). "KefF, the regulatory subunit of the potassium efflux system KefC, shows quinone oxidoreductase activity." J Bacteriol 193(18);4925-32. PMID: 21742892

Miller00a: Miller S, Ness LS, Wood CM, Fox BC, Booth IR (2000). "Identification of an ancillary protein, YabF, required for activity of the KefC glutathione-gated potassium efflux system in Escherichia coli." J Bacteriol 182(22);6536-40. PMID: 11053405

Miller97: Miller S, Douglas RM, Carter P, Booth IR (1997). "Mutations in the glutathione-gated KefC K+ efflux system of Escherichia coli that cause constitutive activation." J Biol Chem 272(40);24942-7. PMID: 9312097

Reed03: Reed JL, Vo TD, Schilling CH, Palsson BO (2003). "An expanded genome-scale model of Escherichia coli K-12 (iJR904 GSM/GPR)." Genome Biol 4(9);R54. PMID: 12952533

Roosild09: Roosild TP, Castronovo S, Miller S, Li C, Rasmussen T, Bartlett W, Gunasekera B, Choe S, Booth IR (2009). "KTN (RCK) domains regulate K+ channels and transporters by controlling the dimer-hinge conformation." Structure 17(6);893-903. PMID: 19523906

Roosild10: Roosild TP, Castronovo S, Healy J, Miller S, Pliotas C, Rasmussen T, Bartlett W, Conway SJ, Booth IR (2010). "Mechanism of ligand-gated potassium efflux in bacterial pathogens." Proc Natl Acad Sci U S A 107(46);19784-9. PMID: 21041667

UniProt13: UniProt Consortium (2013). "UniProt version 2013-08 released on 2013-08-01 00:00:00." Database.

UniProt14: UniProt Consortium (2014). "UniProt version 2014-01 released on 2014-01-01 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."


Report Errors or Provide Feedback
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 Sun Nov 23, 2014, BIOCYC14B.