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Protein

5,10-methylenetetrahydrofolate reductase

Gene

metF

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

Functioni

Methylenetetrahydrofolate reductase required to generate the methyl groups necessary for methionine synthetase to convert homocysteine to methionine.1 Publication

Catalytic activityi

5-methyltetrahydrofolate + NAD(P)+ = 5,10-methylenetetrahydrofolate + NAD(P)H.3 Publications

Cofactori

FAD6 Publications

Kineticsi

  1. KM=17 µM for NADPH2 Publications
  2. KM=3.9 µM for 5,10-methylenetetrahydrofolate2 Publications

    pH dependencei

    Optimum pH is 6.3-6.4.2 Publications

    Pathwayi

    Sites

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Active sitei28 – 281Proton donor/acceptor2 Publications
    Binding sitei88 – 881FAD2 Publications
    Binding sitei120 – 1201Substrate
    Binding sitei152 – 1521FAD2 Publications
    Binding sitei183 – 1831Substrate
    Binding sitei219 – 2191Substrate
    Binding sitei223 – 2231Substrate
    Binding sitei275 – 2751Substrate
    Binding sitei279 – 2791Substrate

    Regions

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Nucleotide bindingi28 – 336NAD
    Nucleotide bindingi59 – 624FAD2 Publications
    Nucleotide bindingi59 – 602NAD
    Nucleotide bindingi118 – 1203FAD2 Publications
    Nucleotide bindingi131 – 1322FAD2 Publications
    Nucleotide bindingi156 – 1594FAD2 Publications
    Nucleotide bindingi165 – 1728FAD2 Publications

    GO - Molecular functioni

    • FAD binding Source: EcoCyc
    • methylenetetrahydrofolate reductase (NAD(P)H) activity Source: EcoCyc

    GO - Biological processi

    • methionine biosynthetic process Source: UniProtKB-KW
    • one-carbon metabolic process Source: GO_Central
    • protein homotetramerization Source: EcoCyc
    • tetrahydrofolate biosynthetic process Source: EcoCyc
    • tetrahydrofolate interconversion Source: UniProtKB-UniPathway
    Complete GO annotation...

    Keywords - Molecular functioni

    Oxidoreductase

    Keywords - Biological processi

    Amino-acid biosynthesis, Methionine biosynthesis

    Keywords - Ligandi

    FAD, Flavoprotein, NAD, NADP

    Enzyme and pathway databases

    BioCyciEcoCyc:METHYLENETHFREDUCT-MONOMER.
    ECOL316407:JW3913-MONOMER.
    MetaCyc:METHYLENETHFREDUCT-MONOMER.
    RETL1328306-WGS:GSTH-2918-MONOMER.
    BRENDAi1.5.1.20. 2026.
    UniPathwayiUPA00193.

    Names & Taxonomyi

    Protein namesi
    Recommended name:
    5,10-methylenetetrahydrofolate reductase (EC:1.5.1.20)
    Gene namesi
    Name:metF
    Ordered Locus Names:b3941, JW3913
    OrganismiEscherichia coli (strain K12)
    Taxonomic identifieri83333 [NCBI]
    Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia
    ProteomesiUP000000318 Componenti: Chromosome UP000000625 Componenti: Chromosome

    Organism-specific databases

    EcoGeneiEG10585. metF.

    Subcellular locationi

    GO - Cellular componenti

    Complete GO annotation...

    Pathology & Biotechi

    Mutagenesis

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Mutagenesisi28 – 281E → Q: Abolishes enzyme activity. 2 Publications
    Mutagenesisi120 – 1201D → N: Strongly reduces enzyme activity. Strongly reduces affinity for 5-methyltetrahydrofolate. 1 Publication
    Mutagenesisi177 – 1771A → V: Increases the propensity to lose its essential flavin cofactor. 1 Publication
    Mutagenesisi223 – 2231F → A: Strongly decreases substrate and NADH binding. 1 Publication
    Mutagenesisi223 – 2231F → L: Slightly reduced enzyme activity. 1 Publication

    Chemistry

    DrugBankiDB03147. Flavin adenine dinucleotide.

    PTM / Processingi

    Molecule processing

    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Chaini1 – 2962965,10-methylenetetrahydrofolate reductasePRO_0000190261Add
    BLAST

    Proteomic databases

    PaxDbiP0AEZ1.
    PRIDEiP0AEZ1.

    Expressioni

    Inductioni

    Repressed by methionine.1 Publication

    Gene expression databases

    GenevestigatoriP0AEZ1.

    Interactioni

    Subunit structurei

    Homotetramer.3 Publications

    Protein-protein interaction databases

    DIPiDIP-6848N.
    IntActiP0AEZ1. 4 interactions.
    STRINGi511145.b3941.

    Structurei

    Secondary structure

    1
    296
    Legend: HelixTurnBeta strand
    Show more details
    Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
    Helixi4 – 1714Combined sources
    Turni18 – 214Combined sources
    Beta strandi24 – 296Combined sources
    Helixi35 – 4915Combined sources
    Beta strandi54 – 585Combined sources
    Helixi67 – 8115Combined sources
    Beta strandi85 – 917Combined sources
    Helixi96 – 10813Combined sources
    Beta strandi113 – 1175Combined sources
    Beta strandi123 – 1253Combined sources
    Helixi132 – 14211Combined sources
    Beta strandi146 – 1516Combined sources
    Helixi162 – 17514Combined sources
    Beta strandi179 – 1824Combined sources
    Helixi188 – 20013Combined sources
    Beta strandi207 – 2115Combined sources
    Helixi217 – 22610Combined sources
    Helixi233 – 2397Combined sources
    Helixi246 – 26621Combined sources
    Beta strandi271 – 2755Combined sources
    Helixi281 – 2899Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1B5TX-ray2.50A/B/C21-294[»]
    1ZP3X-ray1.85A/B/C1-296[»]
    1ZP4X-ray1.85A/B/C1-296[»]
    1ZPTX-ray1.95A/B/C1-296[»]
    1ZRQX-ray2.20A/B/C1-296[»]
    2FMNX-ray2.05A/B/C1-296[»]
    2FMOX-ray2.25A/B/C1-296[»]
    3FSTX-ray1.65A/C/E1-296[»]
    3FSUX-ray1.70A/C/E1-296[»]
    ProteinModelPortaliP0AEZ1.
    SMRiP0AEZ1. Positions 21-295.
    ModBaseiSearch...
    MobiDBiSearch...

    Miscellaneous databases

    EvolutionaryTraceiP0AEZ1.

    Family & Domainsi

    Sequence similaritiesi

    Phylogenomic databases

    eggNOGiCOG0685.
    HOGENOMiHOG000246232.
    InParanoidiP0AEZ1.
    KOiK00297.
    OMAiFIRAETG.
    OrthoDBiEOG6D2KVW.
    PhylomeDBiP0AEZ1.

    Family and domain databases

    Gene3Di3.20.20.220. 1 hit.
    InterProiIPR029041. FAD-linked_oxidoreductase-like.
    IPR003171. Mehydrof_redctse.
    IPR004620. MTHF_reductase_bac.
    [Graphical view]
    PfamiPF02219. MTHFR. 1 hit.
    [Graphical view]
    SUPFAMiSSF51730. SSF51730. 1 hit.
    TIGRFAMsiTIGR00676. fadh2. 1 hit.

    Sequencei

    Sequence statusi: Complete.

    P0AEZ1-1 [UniParc]FASTAAdd to basket

    « Hide

            10         20         30         40         50
    MSFFHASQRD ALNQSLAEVQ GQINVSFEFF PPRTSEMEQT LWNSIDRLSS
    60 70 80 90 100
    LKPKFVSVTY GANSGERDRT HSIIKGIKDR TGLEAAPHLT CIDATPDELR
    110 120 130 140 150
    TIARDYWNNG IRHIVALRGD LPPGSGKPEM YASDLVTLLK EVADFDISVA
    160 170 180 190 200
    AYPEVHPEAK SAQADLLNLK RKVDAGANRA ITQFFFDVES YLRFRDRCVS
    210 220 230 240 250
    AGIDVEIIPG ILPVSNFKQA KKFADMTNVR IPAWMAQMFD GLDDDAETRK
    260 270 280 290
    LVGANIAMDM VKILSREGVK DFHFYTLNRA EMSYAICHTL GVRPGL
    Length:296
    Mass (Da):33,103
    Last modified:July 21, 1986 - v1
    Checksum:iB702702D2BE9521E
    GO

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    V01502 Genomic DNA. Translation: CAA24747.1.
    L19201 Genomic DNA. Translation: AAB03073.1.
    U00096 Genomic DNA. Translation: AAC76923.1.
    AP009048 Genomic DNA. Translation: BAE77369.1.
    PIRiA00462. RDECMH.
    RefSeqiNP_418376.1. NC_000913.3.

    Genome annotation databases

    EnsemblBacteriaiAAC76923; AAC76923; b3941.
    BAE77369; BAE77369; BAE77369.
    GeneIDi948432.
    KEGGiecj:Y75_p3246.
    eco:b3941.
    PATRICi32123403. VBIEscCol129921_4062.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    V01502 Genomic DNA. Translation: CAA24747.1.
    L19201 Genomic DNA. Translation: AAB03073.1.
    U00096 Genomic DNA. Translation: AAC76923.1.
    AP009048 Genomic DNA. Translation: BAE77369.1.
    PIRiA00462. RDECMH.
    RefSeqiNP_418376.1. NC_000913.3.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    Links Updated
    EntryMethodResolution (Å)ChainPositionsPDBsum
    1B5TX-ray2.50A/B/C21-294[»]
    1ZP3X-ray1.85A/B/C1-296[»]
    1ZP4X-ray1.85A/B/C1-296[»]
    1ZPTX-ray1.95A/B/C1-296[»]
    1ZRQX-ray2.20A/B/C1-296[»]
    2FMNX-ray2.05A/B/C1-296[»]
    2FMOX-ray2.25A/B/C1-296[»]
    3FSTX-ray1.65A/C/E1-296[»]
    3FSUX-ray1.70A/C/E1-296[»]
    ProteinModelPortaliP0AEZ1.
    SMRiP0AEZ1. Positions 21-295.
    ModBaseiSearch...
    MobiDBiSearch...

    Protein-protein interaction databases

    DIPiDIP-6848N.
    IntActiP0AEZ1. 4 interactions.
    STRINGi511145.b3941.

    Chemistry

    DrugBankiDB03147. Flavin adenine dinucleotide.

    Proteomic databases

    PaxDbiP0AEZ1.
    PRIDEiP0AEZ1.

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Genome annotation databases

    EnsemblBacteriaiAAC76923; AAC76923; b3941.
    BAE77369; BAE77369; BAE77369.
    GeneIDi948432.
    KEGGiecj:Y75_p3246.
    eco:b3941.
    PATRICi32123403. VBIEscCol129921_4062.

    Organism-specific databases

    EchoBASEiEB0580.
    EcoGeneiEG10585. metF.

    Phylogenomic databases

    eggNOGiCOG0685.
    HOGENOMiHOG000246232.
    InParanoidiP0AEZ1.
    KOiK00297.
    OMAiFIRAETG.
    OrthoDBiEOG6D2KVW.
    PhylomeDBiP0AEZ1.

    Enzyme and pathway databases

    UniPathwayiUPA00193.
    BioCyciEcoCyc:METHYLENETHFREDUCT-MONOMER.
    ECOL316407:JW3913-MONOMER.
    MetaCyc:METHYLENETHFREDUCT-MONOMER.
    RETL1328306-WGS:GSTH-2918-MONOMER.
    BRENDAi1.5.1.20. 2026.

    Miscellaneous databases

    EvolutionaryTraceiP0AEZ1.
    PROiP0AEZ1.

    Gene expression databases

    GenevestigatoriP0AEZ1.

    Family and domain databases

    Gene3Di3.20.20.220. 1 hit.
    InterProiIPR029041. FAD-linked_oxidoreductase-like.
    IPR003171. Mehydrof_redctse.
    IPR004620. MTHF_reductase_bac.
    [Graphical view]
    PfamiPF02219. MTHFR. 1 hit.
    [Graphical view]
    SUPFAMiSSF51730. SSF51730. 1 hit.
    TIGRFAMsiTIGR00676. fadh2. 1 hit.
    ProtoNetiSearch...

    Publicationsi

    « Hide 'large scale' publications
    1. "Nucleotide sequence of metF, the E. coli structural gene for 5-10 methylene tetrahydrofolate reductase and of its control region."
      Saint-Girons I., Duchange N., Zakin M.M., Park I., Margarita D., Ferrara P., Cohen G.N.
      Nucleic Acids Res. 11:6723-6732(1983) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
      Strain: K12.
    2. "Analysis of the Escherichia coli genome. III. DNA sequence of the region from 87.2 to 89.2 minutes."
      Plunkett G. III, Burland V., Daniels D.L., Blattner F.R.
      Nucleic Acids Res. 21:3391-3398(1993) [PubMed] [Europe PMC] [Abstract]
      Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    3. Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
      Strain: K12 / MG1655 / ATCC 47076.
    4. "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.
    5. "Enzymatic synthesis of the methyl group of methionine. 8. Repression-derepression, purification, and properties of 5,10-methylenetetrahydrofolate reductase from Escherichia coli."
      Katzen H.M., Buchanan J.M.
      J. Biol. Chem. 240:825-835(1965) [PubMed] [Europe PMC] [Abstract]
      Cited for: INDUCTION, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR.
    6. "Construction and physical mapping of plasmids containing the metJBLF gene cluster of E. coli K12."
      Zakin M.M., Greene R.C., Dautry-Varsat A., Duchange N., Ferrara P., Py M.C., Margarita D., Cohen G.N.
      Mol. Gen. Genet. 187:101-106(1982) [PubMed] [Europe PMC] [Abstract]
      Cited for: IDENTIFICATION.
    7. "Folate activation and catalysis in methylenetetrahydrofolate reductase from Escherichia coli: roles for aspartate 120 and glutamate 28."
      Trimmer E.E., Ballou D.P., Ludwig M.L., Matthews R.G.
      Biochemistry 40:6216-6226(2001) [PubMed] [Europe PMC] [Abstract]
      Cited for: CATALYTIC ACTIVITY, COFACTOR, ACTIVE SITE, MUTAGENESIS OF GLU-28 AND ASP-120.
    8. "The structure and properties of methylenetetrahydrofolate reductase from Escherichia coli suggest how folate ameliorates human hyperhomocysteinemia."
      Guenther B.D., Sheppard C.A., Tran P., Rozen R., Matthews R.G., Ludwig M.L.
      Nat. Struct. Biol. 6:359-365(1999) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF 21-294 IN COMPLEX WITH FAD, BIOPHYSICOCHEMICAL PROPERTIES, COFACTOR, SUBUNIT, MUTAGENESIS OF ALA-177.
    9. "Structures of NADH and CH3-H4folate complexes of Escherichia coli methylenetetrahydrofolate reductase reveal a spartan strategy for a ping-pong reaction."
      Pejchal R., Sargeant R., Ludwig M.L.
      Biochemistry 44:11447-11457(2005) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) OF WILD TYPE AND MUTANT GLN-28 IN COMPLEXES WITH 5-METHYL-5,6,7,8-TETRAHYDROFOLATE; FAD AND NAD, COFACTOR, SUBUNIT, MUTAGENESIS OF GLU-28, ACTIVE SITE.
    10. "Structural perturbations in the Ala --> Val polymorphism of methylenetetrahydrofolate reductase: how binding of folates may protect against inactivation."
      Pejchal R., Campbell E., Guenther B.D., Lennon B.W., Matthews R.G., Ludwig M.L.
      Biochemistry 45:4808-4818(2006) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (2.05 ANGSTROMS) OF MUTANT VAL-177 IN COMPLEX WITH SUBSTRATE ANALOG LY309887 AND FAD, COFACTOR, SUBUNIT.
    11. "Functional role for the conformationally mobile phenylalanine 223 in the reaction of methylenetetrahydrofolate reductase from Escherichia coli."
      Lee M.N., Takawira D., Nikolova A.P., Ballou D.P., Furtado V.C., Phung N.L., Still B.R., Thorstad M.K., Tanner J.J., Trimmer E.E.
      Biochemistry 48:7673-7685(2009) [PubMed] [Europe PMC] [Abstract]
      Cited for: X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) ON MUTANTS GLN-28 AND LEU-223 IN COMPLEXES WITH 5-METHYL-5,6,7,8-TETRAHYDROFOLATE AND FAD, CATALYTIC ACTIVITY, MUTAGENESIS OF PHE-223, COFACTOR.

    Entry informationi

    Entry nameiMETF_ECOLI
    AccessioniPrimary (citable) accession number: P0AEZ1
    Secondary accession number(s): P00394, Q2M8N7
    Entry historyi
    Integrated into UniProtKB/Swiss-Prot: July 21, 1986
    Last sequence update: July 21, 1986
    Last modified: May 27, 2015
    This is version 89 of the entry and version 1 of the sequence. [Complete history]
    Entry statusiReviewed (UniProtKB/Swiss-Prot)
    Annotation programProkaryotic Protein Annotation Program

    Miscellaneousi

    Keywords - Technical termi

    3D-structure, Complete proteome, Reference proteome

    Documents

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

    External Data

    Dasty 3

    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 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.