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P00383 (DYR21_ECOLX) Reviewed, UniProtKB/Swiss-Prot

Last modified April 16, 2014. Version 83. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (2) | 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:
Dihydrofolate reductase type 2

EC=1.5.1.3
Alternative name(s):
Dihydrofolate reductase type II
Encoded onPlasmid R67
OrganismEscherichia coli
Taxonomic identifier562 [NCBI]
Taxonomic lineageBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Protein attributes

Sequence length78 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.

Pathway

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

Subunit structure

Homotetramer. Ref.3 Ref.4 Ref.5

Domain

The active site is situated at the inner surface of a pore formed by the four subunits. Ref.4 Ref.5

Miscellaneous

Type II plasmid-specified enzyme is practically insensitive to trimethoprim and methotrexate.

Biophysicochemical properties

Kinetic parameters:

KM=5.8 µM for dihydrofolate Ref.3

KM=3.0 µM for NADPH

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 7878Dihydrofolate reductase type 2
PRO_0000186435

Regions

Nucleotide binding66 – 694NADP

Sites

Binding site321NADP
Binding site681Substrate; via amide nitrogen

Experimental info

Mutagenesis651S → A: No effect. Ref.3
Mutagenesis671Q → C: Decreases affinity for NADPH and dihydrofolate about 9-fold. Ref.3
Mutagenesis671Q → H: Increases affinity for dihydrofolate 36-fold. Increases affinity for NADPH 110-fold. Ref.3
Mutagenesis681I → L or M: Decreases affinity for dihydrofolate about 5-fold. Decreases affinity for NADPH about 7-fold. Ref.3
Mutagenesis691Y → F: Decreases affinity for dihydrofolate about 9-fold. Decreases affinity for NADPH about 22-fold. Ref.3
Mutagenesis691Y → H: Decreases affinity for dihydrofolate about 9-fold. Decreases affinity for NADPH about 60-fold. Ref.3

Secondary structure

........... 78
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
P00383 [UniParc].

Last modified July 21, 1986. Version 1.
Checksum: 0BDB0B9146529417

FASTA788,446
        10         20         30         40         50         60 
MERSSNEVSN PVAGNFVFPS NATFGMGDRV RKKSGAAWQG QIVGWYCTNL TPEGYAVESE 

        70 
AHPGSVQIYP VAALERIN 

« Hide

References

[1]"Nucleotide sequence of the dihydrofolate-reductase gene borne by the plasmid R67 and conferring methotrexate resistance."
Brisson N., Hohn T.
Gene 28:271-275(1984) [PubMed] [Europe PMC] [Abstract]
Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA].
[2]"The amino acid sequence of the trimethoprim-resistant dihydrofolate reductase specified in Escherichia coli by R-plasmid R67."
Stone D., Smith S.L.
J. Biol. Chem. 254:10857-10861(1979) [PubMed] [Europe PMC] [Abstract]
Cited for: PROTEIN SEQUENCE.
[3]"Role of S65, Q67, I68, and Y69 residues in homotetrameric R67 dihydrofolate reductase."
Strader M.B., Smiley R.D., Stinnett L.G., VerBerkmoes N.C., Howell E.E.
Biochemistry 40:11344-11352(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, IDENTIFICATION BY MASS SPECTROMETRY, MUTAGENESIS OF SER-65; GLN-67; ILE-68 AND TYR-69.
[4]"A plasmid-encoded dihydrofolate reductase from trimethoprim-resistant bacteria has a novel D2-symmetric active site."
Narayana N., Matthews D.A., Howell E.E., Nguyen-Huu X.
Nat. Struct. Biol. 2:1018-1025(1995) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 17-78 IN COMPLEX WITH FOLATE, DOMAIN, SUBUNIT.
[5]"High-resolution structure of a plasmid-encoded dihydrofolate reductase: pentagonal network of water molecules in the D2-symmetric active site."
Narayana N.
Acta Crystallogr. D 62:695-706(2006) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.1 ANGSTROMS) OF 17-78 OF APOPROTEIN, DOMAIN, SUBUNIT.
[6]"Crystal structure of a type II dihydrofolate reductase catalytic ternary complex."
Krahn J.M., Jackson M.R., DeRose E.F., Howell E.E., London R.E.
Biochemistry 46:14878-14888(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (0.96 ANGSTROMS) OF 17-78 IN COMPLEX WITH DIHYDROFOLATE AND NADP.
[7]"Structure of the Q67H mutant of R67 dihydrofolate reductase-NADP+ complex reveals a novel cofactor binding mode."
Divya N., Grifith E., Narayana N.
Protein Sci. 16:1063-1068(2007) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.15 ANGSTROMS) OF 17-78 OF MUTANT HIS-67 IN COMPLEX WITH NADP.
+Additional computationally mapped references.

Cross-references

Sequence databases

EMBL
GenBank
DDBJ
K02118 Genomic DNA. Translation: AAA26083.1.
PIRRDECD6. A91512.

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1VIEX-ray1.70A17-78[»]
1VIFX-ray1.80A17-78[»]
2GQVX-ray1.10A17-78[»]
2P4TX-ray1.15A17-78[»]
2RH2X-ray0.96A17-78[»]
2RK1X-ray1.26A17-78[»]
2RK2X-ray1.90A17-78[»]
3SFMX-ray1.40A17-78[»]
ProteinModelPortalP00383.
SMRP00383. Positions 19-78.
ModBaseSearch...
MobiDBSearch...

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Enzyme and pathway databases

SABIO-RKP00383.
UniPathwayUPA00077; UER00158.

Family and domain databases

InterProIPR009159. Dhfr_type_II.
IPR008990. Elect_transpt_acc-like_dom.
[Graphical view]
PfamPF06442. DHFR_2. 1 hit.
[Graphical view]
PIRSFPIRSF000199. Dhfr_type_II. 1 hit.
SUPFAMSSF50090. SSF50090. 1 hit.
ProtoNetSearch...

Other

EvolutionaryTraceP00383.

Entry information

Entry nameDYR21_ECOLX
AccessionPrimary (citable) accession number: P00383
Entry history
Integrated into UniProtKB/Swiss-Prot: July 21, 1986
Last sequence update: July 21, 1986
Last modified: April 16, 2014
This is version 83 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Relevant documents

PDB cross-references

Index of Protein Data Bank (PDB) cross-references

PATHWAY comments

Index of metabolic and biosynthesis pathways