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P0ABQ4 (DYR_ECOLI) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 90. 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·Interactions·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Dihydrofolate reductase

EC=1.5.1.3
Gene names
Name:folA
Synonyms:tmrA
Ordered Locus Names:b0048, JW0047
OrganismEscherichia coli (strain K12) [Reference proteome] [HAMAP]
Taxonomic identifier83333 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length159 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Key enzyme in folate metabolism. Catalyzes an essential reaction for de novo glycine and purine synthesis, and for DNA precursor synthesis.

Catalytic activity

5,6,7,8-tetrahydrofolate + NADP+ = 7,8-dihydrofolate + NADPH. Ref.16

Pathway

Cofactor biosynthesis; tetrahydrofolate biosynthesis; 5,6,7,8-tetrahydrofolate from 7,8-dihydrofolate: step 1/1.

Miscellaneous

The strain K12 sequence is shown.

Strain B [RT500] is resistant to 500 micrograms per milliliter of trimethoprim.

Strain B [MB1428] is methotrexate-resistant.

Sequence similarities

Belongs to the dihydrofolate reductase family.

Contains 1 DHFR (dihydrofolate reductase) domain.

Binary interactions

With

Entry

#Exp.

IntAct

Notes

flxAP776094EBI-550404,EBI-553024

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 159159Dihydrofolate reductase
PRO_0000186387

Regions

Domain1 – 158158DHFR
Nucleotide binding13 – 197NADP
Nucleotide binding45 – 462NADP
Nucleotide binding63 – 642NADP
Nucleotide binding95 – 1028NADP

Sites

Binding site51Substrate; via carbonyl oxygen
Binding site71NADP; via amide nitrogen and carbonyl oxygen
Binding site271Substrate
Binding site521Substrate
Binding site571Substrate
Binding site761NADP; via carbonyl oxygen
Binding site1131Substrate

Natural variations

Natural variant281L → R in strain: B[RT500] isozyme 2.
Natural variant301W → G in strain: 1810.
Natural variant1541E → K in strain: B[MB1428].
Natural variant1541E → Q in strain: 1810.

Experimental info

Mutagenesis161M → F or S: Increases catalytic rate about 2-fold. Ref.16
Mutagenesis161M → N: Increases catalytic rate about 2-fold. Increases catalytic rate about 7-fold; when associated with L-20; Y-42; F-92; A-85 and S-152. Ref.16
Mutagenesis201M → I or V: Increases catalytic rate 2-fold. Ref.16
Mutagenesis201M → L: Increases catalytic rate 2.5-fold. Increases catalytic rate about 7-fold; when associated with N-16; Y-42; F-92; A-85 and S-152. Ref.16
Mutagenesis421M → V: Increases catalytic rate almost 2-fold. Ref.16
Mutagenesis421M → Y: Increases catalytic rate almost 2-fold. Increases catalytic rate about 7-fold; when associated with N-16; L-20; A-85; F-92 and S-152. Ref.16
Mutagenesis851C → A: Decreases catalytic rate by one third. Increases catalytic rate about 7-fold; when associated with N-16; L-20; Y-42; F-92 and S-152. Ref.16
Mutagenesis921M → F: No effect. Increases catalytic rate about 7-fold; when associated with N-16; L-20; Y-42; A-85 and S-152. Ref.16
Mutagenesis921M → L: No effect. Ref.16
Mutagenesis1521C → S: Increases catalytic rate 1.5-fold. Increases catalytic rate about 7-fold; when associated with N-16; L-20; Y-42; A-85 and F-92. Ref.16

Secondary structure

.................................. 159
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P0ABQ4 [UniParc].

Last modified July 21, 1986. Version 1.
Checksum: 6A03CDCD7F5F8562

FASTA15917,999
        10         20         30         40         50         60 
MISLIAALAV DRVIGMENAM PWNLPADLAW FKRNTLNKPV IMGRHTWESI GRPLPGRKNI 

        70         80         90        100        110        120 
ILSSQPGTDD RVTWVKSVDE AIAACGDVPE IMVIGGGRVY EQFLPKAQKL YLTHIDAEVE 

       130        140        150 
GDTHFPDYEP DDWESVFSEF HDADAQNSHS YCFEILERR 

« Hide

References

« Hide 'large scale' references
[1]"The amino-acid sequence of the dihydrofolate reductase of a trimethoprim-resistant strain of Escherichia coli."
Stone D., Phillips A.W., Burchall J.J.
Eur. J. Biochem. 72:613-624(1977) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE (ISOZYME 1).
Strain: B [RT500].
[2]"Dihydrofolate reductase: the amino acid sequence of the enzyme from a methotrexate-resistant mutant of Escherichia coli."
Bennett C.D., Rodkey J.A., Sondey J.M., Hirschmann R.
Biochemistry 17:1328-1337(1978) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE.
Strain: B [MB1428].
[3]"Nucleotide sequence of the E coli gene coding for dihydrofolate reductase."
Smith D.R., Calvo J.M.
Nucleic Acids Res. 8:2255-2274(1980) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
Strain: K12.
[4]"Effect of a single amino acid substitution on Escherichia coli dihydrofolate reductase catalysis and ligand binding."
Baccanari D.P., Stone D., Kuyper L.
J. Biol. Chem. 256:1738-1747(1981) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE (ISOZYME 2).
Strain: B [RT500].
[5]"Massive overproduction of dihydrofolate reductase in bacteria as a response to the use of trimethoprim."
Flensburg J., Skoeld O.
Eur. J. Biochem. 162:473-476(1987) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
Strain: 1810.
[6]"Systematic sequencing of the Escherichia coli genome: analysis of the 0-2.4 min region."
Yura T., Mori H., Nagai H., Nagata T., Ishihama A., Fujita N., Isono K., Mizobuchi K., Nakata A.
Nucleic Acids Res. 20:3305-3308(1992) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
Strain: K12.
[7]"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.
[8]"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.
[9]"Escherichia coli proteome analysis using the gene-protein database."
VanBogelen R.A., Abshire K.Z., Moldover B., Olson E.R., Neidhardt F.C.
Electrophoresis 18:1243-1251(1997) [PubMed] [Europe PMC] [Abstract]
Cited for: IDENTIFICATION BY 2D-GEL.
[10]"Crystal structures of Escherichia coli and Lactobacillus casei dihydrofolate reductase refined at 1.7-A resolution. II. Environment of bound NADPH and implications for catalysis."
Filman D.J., Bolin J.T., Matthews D.A., Kraut J.
J. Biol. Chem. 257:13663-13672(1982) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS).
[11]"Crystal structures of Escherichia coli dihydrofolate reductase: the NADP+ holoenzyme and the folate.NADP+ ternary complex. Substrate binding and a model for the transition state."
Bystroff C., Oatley S.J., Kraut J.
Biochemistry 29:3263-3277(1990) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS).
[12]"Crystal structure of unliganded Escherichia coli dihydrofolate reductase. Ligand-induced conformational changes and cooperativity in binding."
Bystroff C., Kraut J.
Biochemistry 30:2227-2239(1991) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS).
[13]"Isomorphous crystal structures of Escherichia coli dihydrofolate reductase complexed with folate, 5-deazafolate, and 5,10-dideazatetrahydrofolate: mechanistic implications."
Reyes V.M., Sawaya M.R., Brown K.A., Kraut J.
Biochemistry 34:2710-2723(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEXES WITH NADPH; FOLATE; 5-DEAZAFOLATE AND 5,10-DIDEAZATETRAHYDROFOLATE.
[14]"Crystal structures of Escherichia coli dihydrofolate reductase complexed with 5-formyltetrahydrofolate (folinic acid) in two space groups: evidence for enolization of pteridine O4."
Lee H., Reyes V.M., Kraut J.
Biochemistry 35:7012-7020(1996) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH 5-FORMYLTETRAHYDROFOLATE.
[15]"Loop and subdomain movements in the mechanism of Escherichia coli dihydrofolate reductase: crystallographic evidence."
Sawaya M.R., Kraut J.
Biochemistry 36:586-603(1997) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.55 ANGSTROMS) IN COMPLEXES WITH NADPH; METHOTREXATE AND TETRAHYDROFOLATE.
[16]"Evolutional design of a hyperactive cysteine- and methionine-free mutant of Escherichia coli dihydrofolate reductase."
Iwakura M., Maki K., Takahashi H., Takenawa T., Yokota A., Katayanagi K., Kamiyama T., Gekko K.
J. Biol. Chem. 281:13234-13246(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) IN COMPLEX WITH FOLATE, CATALYTIC ACTIVITY, MUTAGENESIS OF MET-16; MET-20; MET-42; CYS-85; MET-92 AND CYS-152, IDENTIFICATION BY MASS SPECTROMETRY.
[17]"A 2.13 A structure of E. coli dihydrofolate reductase bound to a novel competitive inhibitor reveals a new binding surface involving the M20 loop region."
Summerfield R.L., Daigle D.M., Mayer S., Mallik D., Hughes D.W., Jackson S.G., Sulek M., Organ M.G., Brown E.D., Junop M.S.
J. Med. Chem. 49:6977-6986(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (2.13 ANGSTROMS).
[18]"Neutron diffraction studies of Escherichia coli dihydrofolate reductase complexed with methotrexate."
Bennett B., Langan P., Coates L., Mustyakimov M., Schoenborn B., Howell E.E., Dealwis C.
Proc. Natl. Acad. Sci. U.S.A. 103:18493-18498(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: STRUCTURE BY NEUTRON DIFFRACTION (2.20 ANGSTROMS) IN COMPLEX WITH METHOTREXATE.
[19]"X-ray structure of the ternary MTX.NADPH complex of the anthrax dihydrofolate reductase: a pharmacophore for dual-site inhibitor design."
Bennett B.C., Wan Q., Ahmad M.F., Langan P., Dealwis C.G.
J. Struct. Biol. 166:162-171(2009) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) IN COMPLEX WITH METHOTREXATE AND NADPH.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
J01609 Genomic DNA. Translation: AAA87976.1.
X05108 Genomic DNA. Translation: CAA28755.1.
U00096 Genomic DNA. Translation: AAC73159.1.
AP009048 Genomic DNA. Translation: BAB96616.1.
PIRRDECD. A93704.
RefSeqNP_414590.1. NC_000913.3.
YP_488354.1. NC_007779.1.

3D structure databases

PDBe
RCSB PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1DDRX-ray2.45A/B1-159[»]
1DDSX-ray2.20A/B1-159[»]
1DHIX-ray1.90A/B1-159[»]
1DHJX-ray1.80A/B1-159[»]
1DRAX-ray1.90A/B1-159[»]
1DRBX-ray1.96A/B1-159[»]
1DREX-ray2.60A1-159[»]
1DRHX-ray2.30A1-159[»]
1DYHX-ray1.90A/B1-159[»]
1DYIX-ray1.90A/B1-159[»]
1DYJX-ray1.85A/B1-159[»]
1JOLX-ray1.96A/B1-159[»]
1JOMX-ray1.90A1-159[»]
1RA1X-ray1.90A1-159[»]
1RA2X-ray1.60A1-159[»]
1RA3X-ray1.80A1-159[»]
1RA8X-ray1.80A1-159[»]
1RA9X-ray1.55A1-159[»]
1RB2X-ray2.10A/B1-159[»]
1RB3X-ray2.30A/B1-159[»]
1RC4X-ray1.90A1-159[»]
1RD7X-ray2.60A/B1-159[»]
1RE7X-ray2.60A/B1-159[»]
1RF7X-ray1.80A1-159[»]
1RG7X-ray2.00A1-159[»]
1RH3X-ray2.40A1-159[»]
1RX1X-ray2.00A1-159[»]
1RX2X-ray1.80A1-159[»]
1RX3X-ray2.20A1-159[»]
1RX4X-ray2.20A1-159[»]
1RX5X-ray2.30A1-159[»]
1RX6X-ray2.00A1-159[»]
1RX7X-ray2.30A1-159[»]
1RX8X-ray2.80A1-159[»]
1RX9X-ray1.90A1-159[»]
1TDRX-ray2.50A/B1-159[»]
2ANOX-ray2.68A1-159[»]
2ANQX-ray2.13A1-159[»]
2D0KX-ray1.90A/B2-159[»]
2DRCX-ray1.90A/B1-159[»]
2INQneutron diffraction2.20A/B1-159[»]
3DAUX-ray1.50A1-159[»]
3DRCX-ray1.90A/B1-159[»]
3K74X-ray1.95A1-159[»]
3KFYX-ray2.08A1-159[»]
3OCHX-ray1.79A/B1-159[»]
3QL3X-ray1.80A1-159[»]
3QYLX-ray1.79A1-159[»]
3QYOX-ray2.09A1-159[»]
3R33X-ray2.09A1-159[»]
4DFRX-ray1.70A/B1-159[»]
4EIGX-ray2.50A1-159[»]
4EIZX-ray2.20A/B1-159[»]
4EJ1X-ray1.75A/B1-159[»]
4FHBX-ray2.80A1-159[»]
4GH8X-ray1.85A/B1-158[»]
4I13X-ray1.60A1-159[»]
4I1NX-ray1.89A1-159[»]
4KJJX-ray1.15A1-159[»]
4KJKX-ray1.35A1-159[»]
4KJLX-ray1.38A1-23[»]
4NX6X-ray1.35A1-159[»]
4NX7X-ray1.15A1-159[»]
5DFRX-ray2.30A1-159[»]
6DFRX-ray2.40A1-159[»]
7DFRX-ray2.50A1-159[»]
DisProtDP00301.
ProteinModelPortalP0ABQ4.
SMRP0ABQ4. Positions 1-159.
ModBaseSearch...
MobiDBSearch...

Protein-protein interaction databases

DIPDIP-35824N.
IntActP0ABQ4. 16 interactions.
MINTMINT-1239602.
STRING511145.b0048.

Chemistry

BindingDBP0ABQ4.
ChEMBLCHEMBL1809.

2D gel databases

SWISS-2DPAGEP0ABQ4.

Proteomic databases

PaxDbP0ABQ4.
PRIDEP0ABQ4.

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Genome annotation databases

EnsemblBacteriaAAC73159; AAC73159; b0048.
BAB96616; BAB96616; BAB96616.
GeneID12933205.
944790.
KEGGecj:Y75_p0048.
eco:b0048.
PATRIC32115195. VBIEscCol129921_0049.

Organism-specific databases

EchoBASEEB0322.
EcoGeneEG10326. folA.

Phylogenomic databases

eggNOGCOG0262.
HOGENOMHOG000040233.
KOK00287.
OMAPTRFVVY.
OrthoDBEOG6KT2V2.
PhylomeDBP0ABQ4.
ProtClustDBPRK10769.

Enzyme and pathway databases

BioCycEcoCyc:DIHYDROFOLATEREDUCT-MONOMER.
ECOL316407:JW0047-MONOMER.
MetaCyc:DIHYDROFOLATEREDUCT-MONOMER.
SABIO-RKP0ABQ4.
UniPathwayUPA00077; UER00158.

Gene expression databases

GenevestigatorP0ABQ4.

Family and domain databases

Gene3D3.40.430.10. 1 hit.
InterProIPR012259. DHFR.
IPR024072. DHFR-like_dom.
IPR017925. DHFR_CS.
IPR001796. DHFR_dom.
[Graphical view]
PfamPF00186. DHFR_1. 1 hit.
[Graphical view]
PIRSFPIRSF000194. DHFR. 1 hit.
PRINTSPR00070. DHFR.
SUPFAMSSF53597. SSF53597. 1 hit.
PROSITEPS00075. DHFR_1. 1 hit.
PS51330. DHFR_2. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceP0ABQ4.
PROP0ABQ4.

Entry information

Entry nameDYR_ECOLI
AccessionPrimary (citable) accession number: P0ABQ4
Secondary accession number(s): P00379
Entry history
Integrated into UniProtKB/Swiss-Prot: July 21, 1986
Last sequence update: July 21, 1986
Last modified: April 16, 2014
This is version 90 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