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P00383

- DYR21_ECOLX

UniProt

P00383 - DYR21_ECOLX

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Protein

Dihydrofolate reductase type 2

Gene
N/A
Organism
Escherichia coli
Status
Reviewed - Annotation score: 5 out of 5- Experimental evidence at protein leveli

Functioni

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

Catalytic activityi

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

Kineticsi

  1. KM=5.8 µM for dihydrofolate1 Publication
  2. KM=3.0 µM for NADPH1 Publication

Pathwayi

Sites

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Binding sitei32 – 321NADP2 Publications
Binding sitei68 – 681Substrate; via amide nitrogen

Regions

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Nucleotide bindingi66 – 694NADP2 Publications

GO - Molecular functioni

  1. dihydrofolate reductase activity Source: UniProtKB-EC

GO - Biological processi

  1. one-carbon metabolic process Source: UniProtKB-KW
  2. response to antibiotic Source: UniProtKB-KW
  3. response to drug Source: InterPro
  4. response to methotrexate Source: UniProtKB-KW
  5. tetrahydrofolate biosynthetic process Source: UniProtKB-UniPathway
Complete GO annotation...

Keywords - Molecular functioni

Oxidoreductase

Keywords - Biological processi

Antibiotic resistance, Methotrexate resistance, One-carbon metabolism, Trimethoprim resistance

Keywords - Ligandi

NADP

Enzyme and pathway databases

SABIO-RKP00383.
UniPathwayiUPA00077; UER00158.

Names & Taxonomyi

Protein namesi
Recommended name:
Dihydrofolate reductase type 2 (EC:1.5.1.3)
Alternative name(s):
Dihydrofolate reductase type II
Encoded oniPlasmid R670 Publication
OrganismiEscherichia coli
Taxonomic identifieri562 [NCBI]
Taxonomic lineageiBacteriaProteobacteriaGammaproteobacteriaEnterobacterialesEnterobacteriaceaeEscherichia

Pathology & Biotechi

Mutagenesis

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Mutagenesisi65 – 651S → A: No effect. 1 Publication
Mutagenesisi67 – 671Q → C: Decreases affinity for NADPH and dihydrofolate about 9-fold. 1 Publication
Mutagenesisi67 – 671Q → H: Increases affinity for dihydrofolate 36-fold. Increases affinity for NADPH 110-fold. 1 Publication
Mutagenesisi68 – 681I → L or M: Decreases affinity for dihydrofolate about 5-fold. Decreases affinity for NADPH about 7-fold. 1 Publication
Mutagenesisi69 – 691Y → F: Decreases affinity for dihydrofolate about 9-fold. Decreases affinity for NADPH about 22-fold. 1 Publication
Mutagenesisi69 – 691Y → H: Decreases affinity for dihydrofolate about 9-fold. Decreases affinity for NADPH about 60-fold. 1 Publication

PTM / Processingi

Molecule processing

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Chaini1 – 7878Dihydrofolate reductase type 2PRO_0000186435Add
BLAST

Interactioni

Subunit structurei

Homotetramer.5 Publications

Structurei

Secondary structure

1
78
Legend: HelixTurnBeta strand
Show more details
Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifierActions
Beta strandi29 – 368
Beta strandi39 – 468
Beta strandi52 – 6211
Beta strandi66 – 705
Helixi71 – 733
Beta strandi74 – 763

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
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[»]
ProteinModelPortaliP00383.
SMRiP00383. Positions 19-78.
ModBaseiSearch...
MobiDBiSearch...

Miscellaneous databases

EvolutionaryTraceiP00383.

Family & Domainsi

Domaini

The active site is situated at the inner surface of a pore formed by the four subunits.2 Publications

Family and domain databases

InterProiIPR009159. Dhfr_type_II.
IPR008990. Elect_transpt_acc-like_dom.
[Graphical view]
PfamiPF06442. DHFR_2. 1 hit.
[Graphical view]
PIRSFiPIRSF000199. Dhfr_type_II. 1 hit.
SUPFAMiSSF50090. SSF50090. 1 hit.

Sequencei

Sequence statusi: Complete.

P00383-1 [UniParc]FASTAAdd to Basket

« Hide

        10         20         30         40         50
MERSSNEVSN PVAGNFVFPS NATFGMGDRV RKKSGAAWQG QIVGWYCTNL
60 70
TPEGYAVESE AHPGSVQIYP VAALERIN
Length:78
Mass (Da):8,446
Last modified:July 21, 1986 - v1
Checksum:i0BDB0B9146529417
GO

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
K02118 Genomic DNA. Translation: AAA26083.1.
PIRiA91512. RDECD6.

Cross-referencesi

Sequence databases

Select the link destinations:
EMBL
GenBank
DDBJ
Links Updated
K02118 Genomic DNA. Translation: AAA26083.1 .
PIRi A91512. RDECD6.

3D structure databases

Select the link destinations:
PDBe
RCSB PDB
PDBj
Links Updated
Entry Method Resolution (Å) Chain Positions PDBsum
1VIE X-ray 1.70 A 17-78 [» ]
1VIF X-ray 1.80 A 17-78 [» ]
2GQV X-ray 1.10 A 17-78 [» ]
2P4T X-ray 1.15 A 17-78 [» ]
2RH2 X-ray 0.96 A 17-78 [» ]
2RK1 X-ray 1.26 A 17-78 [» ]
2RK2 X-ray 1.90 A 17-78 [» ]
3SFM X-ray 1.40 A 17-78 [» ]
ProteinModelPortali P00383.
SMRi P00383. Positions 19-78.
ModBasei Search...
MobiDBi Search...

Protocols and materials databases

Structural Biology Knowledgebase Search...

Enzyme and pathway databases

UniPathwayi UPA00077 ; UER00158 .
SABIO-RK P00383.

Miscellaneous databases

EvolutionaryTracei P00383.

Family and domain databases

InterProi IPR009159. Dhfr_type_II.
IPR008990. Elect_transpt_acc-like_dom.
[Graphical view ]
Pfami PF06442. DHFR_2. 1 hit.
[Graphical view ]
PIRSFi PIRSF000199. Dhfr_type_II. 1 hit.
SUPFAMi SSF50090. SSF50090. 1 hit.
ProtoNeti Search...

Publicationsi

  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.

Entry informationi

Entry nameiDYR21_ECOLX
AccessioniPrimary (citable) accession number: P00383
Entry historyi
Integrated into UniProtKB/Swiss-Prot: July 21, 1986
Last sequence update: July 21, 1986
Last modified: October 1, 2014
This is version 84 of the entry and version 1 of the sequence. [Complete history]
Entry statusiReviewed (UniProtKB/Swiss-Prot)
Annotation programProkaryotic Protein Annotation Program

Miscellaneousi

Miscellaneous

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

Keywords - Technical termi

3D-structure, Direct protein sequencing, Plasmid

Documents

  1. PATHWAY comments
    Index of metabolic and biosynthesis pathways
  2. PDB cross-references
    Index of Protein Data Bank (PDB) cross-references

External Data

Dasty 3