Skip Header

You are using a version of Internet Explorer that may not display all features of this website. Please upgrade to a modern browser.
Contribute Send feedback
Read comments (?) or add your own

P0AEZ1 (METF_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified July 9, 2014. Version 82. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (4) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
5,10-methylenetetrahydrofolate reductase

EC=1.5.1.20
Gene names
Name:metF
Ordered Locus Names:b3941, JW3913
OrganismEscherichia coli (strain K12) [Reference proteome] [HAMAP]
Taxonomic identifier83333 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length296 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Methylenetetrahydrofolate reductase required to generate the methyl groups necessary for methionine synthetase to convert homocysteine to methionine. Ref.5

Catalytic activity

5-methyltetrahydrofolate + NAD(P)+ = 5,10-methylenetetrahydrofolate + NAD(P)H. Ref.5 Ref.7 Ref.11

Cofactor

FAD. Ref.5 Ref.7 Ref.8 Ref.9 Ref.10 Ref.11

Pathway

One-carbon metabolism; tetrahydrofolate interconversion.

Subunit structure

Homotetramer. Ref.8 Ref.9 Ref.10

Induction

Repressed by methionine. Ref.5

Sequence similarities

Belongs to the methylenetetrahydrofolate reductase family.

Biophysicochemical properties

Kinetic parameters:

KM=17 µM for NADPH Ref.5 Ref.8

KM=3.9 µM for 5,10-methylenetetrahydrofolate

pH dependence:

Optimum pH is 6.3-6.4.

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 2962965,10-methylenetetrahydrofolate reductase
PRO_0000190261

Regions

Nucleotide binding28 – 336NAD
Nucleotide binding59 – 624FAD
Nucleotide binding59 – 602NAD
Nucleotide binding118 – 1203FAD
Nucleotide binding131 – 1322FAD
Nucleotide binding156 – 1594FAD
Nucleotide binding165 – 1728FAD

Sites

Active site281Proton donor/acceptor Ref.7 Ref.9
Binding site881FAD
Binding site1201Substrate
Binding site1521FAD
Binding site1831Substrate
Binding site2191Substrate
Binding site2231Substrate
Binding site2751Substrate
Binding site2791Substrate

Experimental info

Mutagenesis281E → Q: Abolishes enzyme activity. Ref.7 Ref.9
Mutagenesis1201D → N: Strongly reduces enzyme activity. Strongly reduces affinity for 5-methyltetrahydrofolate. Ref.7
Mutagenesis1771A → V: Increases the propensity to lose its essential flavin cofactor. Ref.8
Mutagenesis2231F → A: Strongly decreases substrate and NADH binding. Ref.11
Mutagenesis2231F → L: Slightly reduced enzyme activity. Ref.11

Secondary structure

.......................................... 296
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P0AEZ1 [UniParc].

Last modified July 21, 1986. Version 1.
Checksum: B702702D2BE9521E

FASTA29633,103
        10         20         30         40         50         60 
MSFFHASQRD ALNQSLAEVQ GQINVSFEFF PPRTSEMEQT LWNSIDRLSS LKPKFVSVTY 

        70         80         90        100        110        120 
GANSGERDRT HSIIKGIKDR TGLEAAPHLT CIDATPDELR TIARDYWNNG IRHIVALRGD 

       130        140        150        160        170        180 
LPPGSGKPEM YASDLVTLLK EVADFDISVA AYPEVHPEAK SAQADLLNLK RKVDAGANRA 

       190        200        210        220        230        240 
ITQFFFDVES YLRFRDRCVS AGIDVEIIPG ILPVSNFKQA KKFADMTNVR IPAWMAQMFD 

       250        260        270        280        290 
GLDDDAETRK LVGANIAMDM VKILSREGVK DFHFYTLNRA EMSYAICHTL GVRPGL 

« Hide

References

« Hide 'large scale' references
[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]"The complete genome sequence of Escherichia coli K-12."
Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V., Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F., Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B., Shao Y.
Science 277:1453-1462(1997) [PubMed] [Europe PMC] [Abstract]
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.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
V01502 Genomic DNA. Translation: CAA24747.1.
L19201 Genomic DNA. Translation: AAB03073.1.
U00096 Genomic DNA. Translation: AAC76923.1.
AP009048 Genomic DNA. Translation: BAE77369.1.
PIRRDECMH. A00462.
RefSeqNP_418376.1. NC_000913.3.
YP_491510.1. NC_007779.1.

3D structure databases

PDBe
RCSB-PDB
PDBj
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[»]
ProteinModelPortalP0AEZ1.
SMRP0AEZ1. Positions 21-295.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

DIPDIP-6848N.
IntActP0AEZ1. 4 interactions.
STRING511145.b3941.

Proteomic databases

PaxDbP0AEZ1.
PRIDEP0AEZ1.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAC76923; AAC76923; b3941.
BAE77369; BAE77369; BAE77369.
GeneID12934069.
948432.
KEGGecj:Y75_p3246.
eco:b3941.
PATRIC32123403. VBIEscCol129921_4062.

Organism-specific databases

EchoBASEEB0580.
EcoGeneEG10585. metF.

Phylogenomic databases

eggNOGCOG0685.
HOGENOMHOG000246232.
KOK00297.
OMAFEFFPTK.
OrthoDBEOG6D2KVW.
PhylomeDBP0AEZ1.

Enzyme and pathway databases

BioCycEcoCyc:METHYLENETHFREDUCT-MONOMER.
ECOL316407:JW3913-MONOMER.
MetaCyc:METHYLENETHFREDUCT-MONOMER.
RETL1328306-WGS:GSTH-2918-MONOMER.
UniPathwayUPA00193.

Gene expression databases

GenevestigatorP0AEZ1.

Family and domain databases

Gene3D3.20.20.220. 1 hit.
InterProIPR029041. FAD-linked_oxidoreductase-like.
IPR003171. Mehydrof_redctse.
IPR004620. MTHF_reductase_bac.
[Graphical view]
PfamPF02219. MTHFR. 1 hit.
[Graphical view]
SUPFAMSSF51730. SSF51730. 1 hit.
TIGRFAMsTIGR00676. fadh2. 1 hit.
ProtoNetSearch...

Other

EvolutionaryTraceP0AEZ1.
PROP0AEZ1.

Entry information

Entry nameMETF_ECOLI
AccessionPrimary (citable) accession number: P0AEZ1
Secondary accession number(s): P00394, Q2M8N7
Entry history
Integrated into UniProtKB/Swiss-Prot: July 21, 1986
Last sequence update: July 21, 1986
Last modified: July 9, 2014
This is version 82 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

SIMILARITY comments

Index of protein domains and families

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways

Escherichia coli

Escherichia coli (strain K12): entries and cross-references to EcoGene