DYR21_ECOLX
ID DYR21_ECOLX Reviewed; 78 AA.
AC P00383;
DT 21-JUL-1986, integrated into UniProtKB/Swiss-Prot.
DT 21-JUL-1986, sequence version 1.
DT 03-AUG-2022, entry version 106.
DE RecName: Full=Dihydrofolate reductase type 2;
DE EC=1.5.1.3;
DE AltName: Full=Dihydrofolate reductase type II;
OS Escherichia coli.
OG Plasmid R67.
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Escherichia.
OX NCBI_TaxID=562;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX PubMed=6735180; DOI=10.1016/0378-1119(84)90266-x;
RA Brisson N., Hohn T.;
RT "Nucleotide sequence of the dihydrofolate-reductase gene borne by the
RT plasmid R67 and conferring methotrexate resistance.";
RL Gene 28:271-275(1984).
RN [2]
RP PROTEIN SEQUENCE.
RX PubMed=387758; DOI=10.1016/s0021-9258(19)86600-0;
RA Stone D., Smith S.L.;
RT "The amino acid sequence of the trimethoprim-resistant dihydrofolate
RT reductase specified in Escherichia coli by R-plasmid R67.";
RL J. Biol. Chem. 254:10857-10861(1979).
RN [3]
RP BIOPHYSICOCHEMICAL PROPERTIES, SUBUNIT, IDENTIFICATION BY MASS
RP SPECTROMETRY, AND MUTAGENESIS OF SER-65; GLN-67; ILE-68 AND TYR-69.
RX PubMed=11560482; DOI=10.1021/bi0110544;
RA Strader M.B., Smiley R.D., Stinnett L.G., VerBerkmoes N.C., Howell E.E.;
RT "Role of S65, Q67, I68, and Y69 residues in homotetrameric R67
RT dihydrofolate reductase.";
RL Biochemistry 40:11344-11352(2001).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (1.7 ANGSTROMS) OF 17-78 IN COMPLEX WITH FOLATE,
RP DOMAIN, AND SUBUNIT.
RX PubMed=7583655; DOI=10.1038/nsb1195-1018;
RA Narayana N., Matthews D.A., Howell E.E., Nguyen-Huu X.;
RT "A plasmid-encoded dihydrofolate reductase from trimethoprim-resistant
RT bacteria has a novel D2-symmetric active site.";
RL Nat. Struct. Biol. 2:1018-1025(1995).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.1 ANGSTROMS) OF 17-78 OF APOPROTEIN, DOMAIN, AND
RP SUBUNIT.
RX PubMed=16790925; DOI=10.1107/s0907444906014764;
RA Narayana N.;
RT "High-resolution structure of a plasmid-encoded dihydrofolate reductase:
RT pentagonal network of water molecules in the D2-symmetric active site.";
RL Acta Crystallogr. D 62:695-706(2006).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (0.96 ANGSTROMS) OF 17-78 IN COMPLEX WITH
RP DIHYDROFOLATE AND NADP.
RX PubMed=18052202; DOI=10.1021/bi701532r;
RA Krahn J.M., Jackson M.R., DeRose E.F., Howell E.E., London R.E.;
RT "Crystal structure of a type II dihydrofolate reductase catalytic ternary
RT complex.";
RL Biochemistry 46:14878-14888(2007).
RN [7]
RP X-RAY CRYSTALLOGRAPHY (1.15 ANGSTROMS) OF 17-78 OF MUTANT HIS-67 IN COMPLEX
RP WITH NADP.
RX PubMed=17473013; DOI=10.1110/ps.062740907;
RA Divya N., Grifith E., Narayana N.;
RT "Structure of the Q67H mutant of R67 dihydrofolate reductase-NADP+ complex
RT reveals a novel cofactor binding mode.";
RL Protein Sci. 16:1063-1068(2007).
CC -!- FUNCTION: Key enzyme in folate metabolism. Catalyzes an essential
CC reaction for de novo glycine and purine synthesis, and for DNA
CC precursor synthesis.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6S)-5,6,7,8-tetrahydrofolate + NADP(+) = 7,8-dihydrofolate +
CC H(+) + NADPH; Xref=Rhea:RHEA:15009, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:57451, ChEBI:CHEBI:57453, ChEBI:CHEBI:57783,
CC ChEBI:CHEBI:58349; EC=1.5.1.3;
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=5.8 uM for dihydrofolate {ECO:0000269|PubMed:11560482};
CC KM=3.0 uM for NADPH {ECO:0000269|PubMed:11560482};
CC -!- PATHWAY: Cofactor biosynthesis; tetrahydrofolate biosynthesis; 5,6,7,8-
CC tetrahydrofolate from 7,8-dihydrofolate: step 1/1.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:11560482,
CC ECO:0000269|PubMed:16790925, ECO:0000269|PubMed:17473013,
CC ECO:0000269|PubMed:18052202, ECO:0000269|PubMed:7583655}.
CC -!- DOMAIN: The active site is situated at the inner surface of a pore
CC formed by the four subunits. {ECO:0000269|PubMed:16790925,
CC ECO:0000269|PubMed:7583655}.
CC -!- MISCELLANEOUS: Type II plasmid-specified enzyme is practically
CC insensitive to trimethoprim and methotrexate.
CC ---------------------------------------------------------------------------
CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms
CC Distributed under the Creative Commons Attribution (CC BY 4.0) License
CC ---------------------------------------------------------------------------
DR EMBL; K02118; AAA26083.1; -; Genomic_DNA.
DR PIR; A91512; RDECD6.
DR RefSeq; WP_000442373.1; NZ_FCPF01000114.1.
DR PDB; 1VIE; X-ray; 1.70 A; A=17-78.
DR PDB; 1VIF; X-ray; 1.80 A; A=17-78.
DR PDB; 2GQV; X-ray; 1.10 A; A=17-78.
DR PDB; 2P4T; X-ray; 1.15 A; A=17-78.
DR PDB; 2RH2; X-ray; 0.96 A; A=17-78.
DR PDB; 2RK1; X-ray; 1.26 A; A=17-78.
DR PDB; 2RK2; X-ray; 1.90 A; A=17-78.
DR PDB; 3SFM; X-ray; 1.40 A; A=17-78.
DR PDB; 6NXZ; X-ray; 1.75 A; A=17-78.
DR PDB; 6NY0; X-ray; 1.40 A; A=18-78.
DR PDBsum; 1VIE; -.
DR PDBsum; 1VIF; -.
DR PDBsum; 2GQV; -.
DR PDBsum; 2P4T; -.
DR PDBsum; 2RH2; -.
DR PDBsum; 2RK1; -.
DR PDBsum; 2RK2; -.
DR PDBsum; 3SFM; -.
DR PDBsum; 6NXZ; -.
DR PDBsum; 6NY0; -.
DR AlphaFoldDB; P00383; -.
DR BMRB; P00383; -.
DR SMR; P00383; -.
DR BRENDA; 1.5.1.3; 2026.
DR SABIO-RK; P00383; -.
DR UniPathway; UPA00077; UER00158.
DR EvolutionaryTrace; P00383; -.
DR GO; GO:0004146; F:dihydrofolate reductase activity; IEA:UniProtKB-EC.
DR GO; GO:0006730; P:one-carbon metabolic process; IEA:UniProtKB-KW.
DR GO; GO:0046677; P:response to antibiotic; IEA:UniProtKB-KW.
DR GO; GO:0031427; P:response to methotrexate; IEA:UniProtKB-KW.
DR GO; GO:0009410; P:response to xenobiotic stimulus; IEA:InterPro.
DR GO; GO:0046654; P:tetrahydrofolate biosynthetic process; IEA:UniProtKB-UniPathway.
DR Gene3D; 2.30.30.60; -; 1.
DR InterPro; IPR009159; Dhfr_type_II.
DR InterPro; IPR008990; Elect_transpt_acc-like_dom_sf.
DR InterPro; IPR023408; MscS_dom_sf.
DR Pfam; PF06442; DHFR_2; 1.
DR PIRSF; PIRSF000199; Dhfr_type_II; 1.
DR SUPFAM; SSF50090; SSF50090; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Antibiotic resistance; Direct protein sequencing;
KW Methotrexate resistance; NADP; One-carbon metabolism; Oxidoreductase;
KW Plasmid; Trimethoprim resistance.
FT CHAIN 1..78
FT /note="Dihydrofolate reductase type 2"
FT /id="PRO_0000186435"
FT BINDING 32..36
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:17473013,
FT ECO:0000269|PubMed:18052202, ECO:0007744|PDB:2P4T,
FT ECO:0007744|PDB:2RK1, ECO:0007744|PDB:2RK2"
FT BINDING 66..69
FT /ligand="NADP(+)"
FT /ligand_id="ChEBI:CHEBI:58349"
FT /evidence="ECO:0000269|PubMed:17473013,
FT ECO:0000269|PubMed:18052202, ECO:0007744|PDB:2P4T,
FT ECO:0007744|PDB:2RK1, ECO:0007744|PDB:2RK2"
FT BINDING 68
FT /ligand="substrate"
FT /evidence="ECO:0000269|PubMed:18052202,
FT ECO:0007744|PDB:2RK1"
FT MUTAGEN 65
FT /note="S->A: No effect."
FT /evidence="ECO:0000269|PubMed:11560482"
FT MUTAGEN 67
FT /note="Q->C: Decreases affinity for NADPH and dihydrofolate
FT about 9-fold."
FT /evidence="ECO:0000269|PubMed:11560482"
FT MUTAGEN 67
FT /note="Q->H: Increases affinity for dihydrofolate 36-fold.
FT Increases affinity for NADPH 110-fold."
FT /evidence="ECO:0000269|PubMed:11560482"
FT MUTAGEN 68
FT /note="I->L,M: Decreases affinity for dihydrofolate about
FT 5-fold. Decreases affinity for NADPH about 7-fold."
FT /evidence="ECO:0000269|PubMed:11560482"
FT MUTAGEN 69
FT /note="Y->F: Decreases affinity for dihydrofolate about 9-
FT fold. Decreases affinity for NADPH about 22-fold."
FT /evidence="ECO:0000269|PubMed:11560482"
FT MUTAGEN 69
FT /note="Y->H: Decreases affinity for dihydrofolate about 9-
FT fold. Decreases affinity for NADPH about 60-fold."
FT /evidence="ECO:0000269|PubMed:11560482"
FT STRAND 29..36
FT /evidence="ECO:0007829|PDB:2RH2"
FT STRAND 39..46
FT /evidence="ECO:0007829|PDB:2RH2"
FT STRAND 52..62
FT /evidence="ECO:0007829|PDB:2RH2"
FT STRAND 66..70
FT /evidence="ECO:0007829|PDB:2RH2"
FT HELIX 71..73
FT /evidence="ECO:0007829|PDB:2RH2"
FT STRAND 74..76
FT /evidence="ECO:0007829|PDB:2RH2"
SQ SEQUENCE 78 AA; 8446 MW; 0BDB0B9146529417 CRC64;
MERSSNEVSN PVAGNFVFPS NATFGMGDRV RKKSGAAWQG QIVGWYCTNL TPEGYAVESE
AHPGSVQIYP VAALERIN
