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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

  1. FAD binding Source: EcoCyc
  2. methylenetetrahydrofolate reductase (NAD(P)H) activity Source: EcoCyc

GO - Biological processi

  1. methionine biosynthetic process Source: UniProtKB-KW
  2. one-carbon metabolic process Source: GO_Central
  3. protein homotetramerization Source: EcoCyc
  4. tetrahydrofolate biosynthetic process Source: EcoCyc
  5. 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

  1. cytosol Source: GO_Central
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.
YP_491510.1. NC_007779.1.

Genome annotation databases

EnsemblBacteriaiAAC76923; AAC76923; b3941.
BAE77369; BAE77369; BAE77369.
GeneIDi12934069.
948432.
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.
YP_491510.1. NC_007779.1.

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.
GeneIDi12934069.
948432.
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: April 1, 2015
This is version 88 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.