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Protein

Dihydrofolate reductase type 2

Gene
N/A
Organism
Escherichia coli
Status
Reviewed-Annotation score: 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

    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

    GO - Biological processi

    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

    BRENDAi1.5.1.3. 2026.
    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 – 368Combined sources
    Beta strandi39 – 468Combined sources
    Beta strandi52 – 6211Combined sources
    Beta strandi66 – 705Combined sources
    Helixi71 – 733Combined sources
    Beta strandi74 – 763Combined sources

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    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:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    K02118 Genomic DNA. Translation: AAA26083.1.
    PIRiA91512. RDECD6.

    Cross-referencesi

    Sequence databases

    Select the link destinations:
    EMBLi
    GenBanki
    DDBJi
    Links Updated
    K02118 Genomic DNA. Translation: AAA26083.1.
    PIRiA91512. RDECD6.

    3D structure databases

    Select the link destinations:
    PDBei
    RCSB PDBi
    PDBji
    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...

    Protocols and materials databases

    Structural Biology KnowledgebaseSearch...

    Enzyme and pathway databases

    UniPathwayiUPA00077; UER00158.
    BRENDAi1.5.1.3. 2026.
    SABIO-RKP00383.

    Miscellaneous databases

    EvolutionaryTraceiP00383.

    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.
    ProtoNetiSearch...

    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: May 27, 2015
    This is version 87 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

    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.