Skip Header

You are using a version of browser that may not display all the features of this website. Please consider upgrading your browser.
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

    Pathway:itetrahydrofolate biosynthesis

    This protein is involved in step 1 of the subpathway that synthesizes 5,6,7,8-tetrahydrofolate from 7,8-dihydrofolate.
    Proteins known to be involved in this subpathway in this organism are:
    1. Dihydrofolate reductase type 8 (dhfrVIII), Dihydrofolate reductase type 9 (dhfrIX), Dihydrofolate reductase (folA), Dihydrofolate reductase (dfrA26), Dihydrofolate reductase (dfrA12), Dihydrofolate reductase (dfrA12), Dihydrofolate reductase type A10 (dfrA10), Dihydrofolate reductase (dfrA21), Dihydrofolate reductase type A13 (dfrA13), Dihydrofolate reductase (ECs0051), Dihydrofolate reductase (folA), Dihydrofolate reductase type 2, Dihydrofolate reductase type 2, Dihydrofolate reductase type 2, Dihydrofolate reductase type 7 (dhfrVII), Dihydrofolate reductase type 1 (dhfrI), Dihydrofolate reductase (dfr22), Dihydrofolate reductase type 5 (dhfrV), Dihydrofolate reductase (dfrA12), Dihydrofolate reductase type 15 (dhfrXV), Dihydrofolate reductase (folA), Dihydrofolate reductase (dfrA12), Dihydrofolate reductase (dfrA12), Dihydrofolate reductase (dfrA12), Dihydrofolate reductase (dfr22), Dihydrofolate reductase (dfrA12)
    This subpathway is part of the pathway tetrahydrofolate biosynthesis, which is itself part of Cofactor biosynthesis.
    View all proteins of this organism that are known to be involved in the subpathway that synthesizes 5,6,7,8-tetrahydrofolate from 7,8-dihydrofolate, the pathway tetrahydrofolate biosynthesis and in Cofactor biosynthesis.

    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.