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P00383

- DYR21_ECOLX

UniProt

P00383 - DYR21_ECOLX

Protein

Dihydrofolate reductase type 2

Gene
N/A
Organism
Escherichia coli
Status
Reviewed - Annotation score: 5 out of 5- Experimental evidence at protein leveli
  1. 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

    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

    MERSSNEVSN PVAGNFVFPS NATFGMGDRV RKKSGAAWQG QIVGWYCTNL   50
    TPEGYAVESE AHPGSVQIYP VAALERIN 78
    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