ID   CODA_ECOLI              Reviewed;         427 AA.
AC   P25524; Q2MC87;
DT   01-MAY-1992, integrated into UniProtKB/Swiss-Prot.
DT   23-JAN-2007, sequence version 3.
DT   27-MAR-2024, entry version 174.
DE   RecName: Full=Cytosine deaminase {ECO:0000303|PubMed:1640834, ECO:0000303|PubMed:8226944};
DE            Short=CD {ECO:0000303|PubMed:15381761};
DE            Short=CDA {ECO:0000303|PubMed:21545144};
DE            Short=CDase {ECO:0000303|PubMed:8226944};
DE            EC=3.5.4.1 {ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:21545144, ECO:0000269|PubMed:21604715, ECO:0000269|PubMed:8226944};
DE   AltName: Full=Cytosine aminohydrolase;
DE   AltName: Full=Isoguanine deaminase {ECO:0000303|PubMed:21604715};
DE            EC=3.5.4.- {ECO:0000269|PubMed:21604715};
GN   Name=codA; OrderedLocusNames=b0337, JW0328;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PROTEIN SEQUENCE OF 2-20.
RC   STRAIN=CSH01;
RX   PubMed=1640834; DOI=10.1111/j.1365-2958.1992.tb00854.x;
RA   Danielsen S., Kilstrup M., Barilla K., Jochimsen B., Neuhard J.;
RT   "Characterization of the Escherichia coli codBA operon encoding cytosine
RT   permease and cytosine deaminase.";
RL   Mol. Microbiol. 6:1335-1344(1992).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RX   PubMed=8450832;
RA   Austin E.A., Huber B.E.;
RT   "A first step in the development of gene therapy for colorectal carcinoma:
RT   cloning, sequencing, and expression of Escherichia coli cytosine
RT   deaminase.";
RL   Mol. Pharmacol. 43:380-387(1993).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RA   Chung E., Allen E., Araujo R., Aparicio A.M., Davis K., Duncan M.,
RA   Federspiel N., Hyman R., Kalman S., Komp C., Kurdi O., Lew H., Lin D.,
RA   Namath A., Oefner P., Roberts D., Schramm S., Davis R.W.;
RT   "Sequence of minutes 4-25 of Escherichia coli.";
RL   Submitted (JAN-1997) to the EMBL/GenBank/DDBJ databases.
RN   [4]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA   Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [5]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT   and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [6]
RP   FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP   ACTIVITY REGULATION.
RX   PubMed=8226944; DOI=10.1016/s0021-9258(20)80485-2;
RA   Porter D.J., Austin E.A.;
RT   "Cytosine deaminase. The roles of divalent metal ions in catalysis.";
RL   J. Biol. Chem. 268:24005-24011(1993).
RN   [7]
RP   SUBUNIT.
RX   PubMed=11679731; DOI=10.1107/s0907444901011064;
RA   Ireton G.C., Black M.E., Stoddard B.L.;
RT   "Crystallization and preliminary X-ray analysis of bacterial cytosine
RT   deaminase.";
RL   Acta Crystallogr. D 57:1643-1645(2001).
RN   [8]
RP   REVIEW, AND BIOTECHNOLOGY.
RX   PubMed=25338741; DOI=10.1208/s12248-014-9675-7;
RA   Zhang J., Kale V., Chen M.;
RT   "Gene-directed enzyme prodrug therapy.";
RL   AAPS J. 17:102-110(2015).
RN   [9]
RP   X-RAY CRYSTALLOGRAPHY (1.75 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH A
RP   MECHANISM-BASED INHIBITOR AND IRON, REACTION MECHANISM, AND ACTIVE SITE.
RX   PubMed=11812140; DOI=10.1006/jmbi.2001.5277;
RA   Ireton G.C., McDermott G., Black M.E., Stoddard B.L.;
RT   "The structure of Escherichia coli cytosine deaminase.";
RL   J. Mol. Biol. 315:687-697(2002).
RN   [10]
RP   X-RAY CRYSTALLOGRAPHY (1.12 ANGSTROMS) OF MUTANTS GLY-314; ALA-314 AND
RP   SER-315 UNCOMPLEXED AND IN COMPLEX WITH
RP   (4S)-5-FLUORO-4-HYDROXY-3,4-DIHYDROPYRIMIDIN-2(1H)-ONE AND IRON, FUNCTION,
RP   CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF ASP-314,
RP   AND BIOTECHNOLOGY.
RX   PubMed=15381761; DOI=10.1093/protein/gzh074;
RA   Mahan S.D., Ireton G.C., Knoeber C., Stoddard B.L., Black M.E.;
RT   "Random mutagenesis and selection of Escherichia coli cytosine deaminase
RT   for cancer gene therapy.";
RL   Protein Eng. Des. Sel. 17:625-633(2004).
RN   [11]
RP   X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) OF 5-427 OF MUTANT
RP   ALA-153/CYS-317/GLY-318 IN COMPLEX WITH IRON, AND BIOTECHNOLOGY.
RX   PubMed=19487291; DOI=10.1158/0008-5472.can-09-0615;
RA   Fuchita M., Ardiani A., Zhao L., Serve K., Stoddard B.L., Black M.E.;
RT   "Bacterial cytosine deaminase mutants created by molecular engineering show
RT   improved 5-fluorocytosine-mediated cell killing in vitro and in vivo.";
RL   Cancer Res. 69:4791-4799(2009).
RN   [12]
RP   X-RAY CRYSTALLOGRAPHY (1.71 ANGSTROMS) IN COMPLEX WITH PHOSPHONOCYTOSINE
RP   INHIBITOR AND ZINC, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, SUBSTRATE
RP   SPECIFICITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION,
RP   MUTAGENESIS OF GLN-157; GLU-218; HIS-247 AND ASP-314, ACTIVE SITES, AND
RP   REACTION MECHANISM.
RC   STRAIN=K12;
RX   PubMed=21545144; DOI=10.1021/bi200483k;
RA   Hall R.S., Fedorov A.A., Xu C., Fedorov E.V., Almo S.C., Raushel F.M.;
RT   "Three-dimensional structure and catalytic mechanism of cytosine
RT   deaminase.";
RL   Biochemistry 50:5077-5085(2011).
RN   [13]
RP   X-RAY CRYSTALLOGRAPHY (2.26 ANGSTROMS) IN COMPLEX WITH ISOGUANINE AND ZINC,
RP   IDENTIFICATION BY MASS SPECTROMETRY, FUNCTION, CATALYTIC ACTIVITY,
RP   IDENTIFICATION OF ISOGUANINE AS SUBSTRATE, BIOPHYSICOCHEMICAL PROPERTIES,
RP   AND DISRUPTION PHENOTYPE.
RX   PubMed=21604715; DOI=10.1021/bi200680y;
RA   Hitchcock D.S., Fedorov A.A., Fedorov E.V., Dangott L.J., Almo S.C.,
RA   Raushel F.M.;
RT   "Rescue of the orphan enzyme isoguanine deaminase.";
RL   Biochemistry 50:5555-5557(2011).
RN   [14]
RP   X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) IN COMPLEX WITH ZINC.
RA   Fedorov A.A., Fedorov E.V., Kamat S., Hitchcock D., Raushel F.M.,
RA   Almo S.C.;
RT   "Crystal structure of cytosine deaminase from Escherichia coli complexed
RT   with two zinc atoms in the active site.";
RL   Submitted (MAR-2011) to the PDB data bank.
CC   -!- FUNCTION: Catalyzes the hydrolytic deamination of cytosine to uracil.
CC       Is involved in the pyrimidine salvage pathway, which allows the cell to
CC       utilize cytosine for pyrimidine nucleotide synthesis. Is also able to
CC       catalyze deamination of isoguanine, a mutagenic oxidation product of
CC       adenine in DNA, and of isocytosine. To a lesser extent, also catalyzes
CC       the conversion of 5-fluorocytosine (5FC) to 5-fluorouracil (5FU); this
CC       activity allows the formation of a cytotoxic chemotherapeutic agent
CC       from a non-cytotoxic precursor. {ECO:0000269|PubMed:15381761,
CC       ECO:0000269|PubMed:21545144, ECO:0000269|PubMed:21604715,
CC       ECO:0000269|PubMed:8226944}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=cytosine + H(+) + H2O = NH4(+) + uracil; Xref=Rhea:RHEA:20605,
CC         ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:16040,
CC         ChEBI:CHEBI:17568, ChEBI:CHEBI:28938; EC=3.5.4.1;
CC         Evidence={ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:21545144,
CC         ECO:0000269|PubMed:21604715, ECO:0000269|PubMed:8226944};
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=H(+) + H2O + isoguanine = NH4(+) + xanthine;
CC         Xref=Rhea:RHEA:47720, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:17712, ChEBI:CHEBI:28938, ChEBI:CHEBI:62462;
CC         Evidence={ECO:0000269|PubMed:21604715};
CC   -!- COFACTOR:
CC       Name=Fe(2+); Xref=ChEBI:CHEBI:29033;
CC         Evidence={ECO:0000269|PubMed:21545144, ECO:0000269|PubMed:8226944};
CC       Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC         Evidence={ECO:0000269|PubMed:8226944};
CC       Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC         Evidence={ECO:0000269|PubMed:21545144, ECO:0000269|PubMed:8226944};
CC       Note=The purified enzyme contains a mixture of Fe(2+) and Zn(2+) bound
CC       in the active site, and a single equivalent of metal is required for
CC       full catalytic activity. After removal of the metal, the reconstitution
CC       of the enzyme with Fe(2+) gives the highest activity, followed by
CC       Mn(2+), and, to a much lesser extent, Co(2+) and Zn(2+).
CC       {ECO:0000269|PubMed:21545144, ECO:0000269|PubMed:8226944};
CC   -!- ACTIVITY REGULATION: Fe(2+)-CDase is rapidly inactivated by H(2)O(2),
CC       whereas Mn(2+)-CDase, Co(2+)-CDase, and Zn(2+)-CDase are not
CC       inactivated by H(2)O(2). CDase is also inhibited by excess divalent
CC       cations (PubMed:8226944). Phosphonocytosine, a mimic of the tetrahedral
CC       reaction intermediate, inhibits the deamination of cytosine with a Ki
CC       of 52 nM (PubMed:21545144). {ECO:0000269|PubMed:21545144,
CC       ECO:0000269|PubMed:8226944}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=0.22 mM for cytosine {ECO:0000269|PubMed:8226944};
CC         KM=0.2 mM for cytosine {ECO:0000269|PubMed:15381761};
CC         KM=3.3 mM for 5-fluorocytosine {ECO:0000269|PubMed:15381761};
CC         KM=0.97 mM for cytosine {ECO:0000269|PubMed:21545144};
CC         KM=0.46 mM for isocytosine {ECO:0000269|PubMed:21545144};
CC         KM=25 mM for creatinine {ECO:0000269|PubMed:21545144};
CC         KM=4.1 mM for 3-oxauracil {ECO:0000269|PubMed:21545144};
CC         KM=72 uM for isoguanine {ECO:0000269|PubMed:21604715};
CC         KM=302 uM for cytosine {ECO:0000269|PubMed:21604715};
CC         Note=kcat is 185 sec(-1) for the deamination of cytosine using Fe(2+)
CC         as cofactor, kcat is 92 sec(-1) using Mn(2+) as cofactor, kcat is 52
CC         sec(-1) using Co(2+) as cofactor, and kcat is 32 sec(-1) using Zn(2+)
CC         as cofactor (PubMed:8226944). kcat is 165 sec(-1) for the deamination
CC         of cytosine and 75.6 sec(-1) for the deamination of 5-fluorocytosine
CC         (PubMed:15381761). kcat is 132 sec(-1) for the deamination of
CC         cytosine, 5.1 sec(-1) for the deamination of isocytosine, 5.6 sec(-1)
CC         for the deamination of creatinine and 2.3 sec(-1) for the hydrolysis
CC         of 3-oxauracil (PubMed:21545144). kcat is 45 sec(-1) for the
CC         deamination of cytosine and 49 sec(-1) for the deamination of
CC         isoguanine at pH 7.7 (PubMed:21604715). {ECO:0000269|PubMed:15381761,
CC         ECO:0000269|PubMed:21545144, ECO:0000269|PubMed:21604715,
CC         ECO:0000269|PubMed:8226944};
CC       pH dependence:
CC         Activity is lost under pH 5 but not affected up to pH 10.
CC         {ECO:0000269|PubMed:21545144};
CC   -!- SUBUNIT: Homohexamer. {ECO:0000269|PubMed:11679731}.
CC   -!- INTERACTION:
CC       P25524; P25524: codA; NbExp=3; IntAct=EBI-559181, EBI-559181;
CC   -!- DISRUPTION PHENOTYPE: Cells lacking this gene have less than 1% of the
CC       isoguanine deaminase activity of the wild-type strain.
CC       {ECO:0000269|PubMed:21604715}.
CC   -!- BIOTECHNOLOGY: Cytosine deaminase is being explored for use as a
CC       suicide gene for cancer gene therapy. The cytosine deaminase/5-
CC       fluorouracil combined therapy has been used successfully for a variety
CC       of animal tumor models and is currently under investigation for the
CC       treatment of human cancers. {ECO:0000269|PubMed:19487291,
CC       ECO:0000303|PubMed:25338741, ECO:0000305|PubMed:15381761}.
CC   -!- SIMILARITY: Belongs to the metallo-dependent hydrolases superfamily.
CC       Cytosine deaminase family. {ECO:0000305}.
CC   -!- SEQUENCE CAUTION:
CC       Sequence=AAB18061.1; Type=Erroneous initiation; Note=Extended N-terminus.; Evidence={ECO:0000305};
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DR   EMBL; X63656; CAA45196.1; -; Genomic_DNA.
DR   EMBL; S56903; AAB25761.2; -; Genomic_DNA.
DR   EMBL; U73857; AAB18061.1; ALT_INIT; Genomic_DNA.
DR   EMBL; U00096; AAC73440.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAE76119.1; -; Genomic_DNA.
DR   PIR; S22662; S22662.
DR   RefSeq; NP_414871.1; NC_000913.3.
DR   RefSeq; WP_001301240.1; NZ_SSZK01000097.1.
DR   PDB; 1K6W; X-ray; 1.75 A; A=2-427.
DR   PDB; 1K70; X-ray; 1.80 A; A=2-427.
DR   PDB; 1R9X; X-ray; 1.58 A; A=2-427.
DR   PDB; 1R9Y; X-ray; 1.57 A; A=2-427.
DR   PDB; 1R9Z; X-ray; 1.32 A; A=2-427.
DR   PDB; 1RA0; X-ray; 1.12 A; A=2-427.
DR   PDB; 1RA5; X-ray; 1.40 A; A=2-427.
DR   PDB; 1RAK; X-ray; 1.32 A; A=2-427.
DR   PDB; 3G77; X-ray; 1.80 A; A=5-427.
DR   PDB; 3O7U; X-ray; 1.71 A; A=1-427.
DR   PDB; 3R0D; X-ray; 1.50 A; A=1-427.
DR   PDB; 3RN6; X-ray; 2.26 A; A=1-427.
DR   PDBsum; 1K6W; -.
DR   PDBsum; 1K70; -.
DR   PDBsum; 1R9X; -.
DR   PDBsum; 1R9Y; -.
DR   PDBsum; 1R9Z; -.
DR   PDBsum; 1RA0; -.
DR   PDBsum; 1RA5; -.
DR   PDBsum; 1RAK; -.
DR   PDBsum; 3G77; -.
DR   PDBsum; 3O7U; -.
DR   PDBsum; 3R0D; -.
DR   PDBsum; 3RN6; -.
DR   AlphaFoldDB; P25524; -.
DR   SMR; P25524; -.
DR   BioGRID; 4259813; 4.
DR   BioGRID; 849391; 1.
DR   DIP; DIP-9306N; -.
DR   IntAct; P25524; 6.
DR   STRING; 511145.b0337; -.
DR   DrugBank; DB03939; (4S)-4-hydroxy-3,4-dihydro-2(1H)-pyrimidinone.
DR   DrugBank; DB04135; (4S)-5-Fluoro-4-hydroxy-3,4-dihydro-2(1H)-pyrimidinone.
DR   jPOST; P25524; -.
DR   PaxDb; 511145-b0337; -.
DR   EnsemblBacteria; AAC73440; AAC73440; b0337.
DR   GeneID; 944996; -.
DR   KEGG; ecj:JW0328; -.
DR   KEGG; eco:b0337; -.
DR   PATRIC; fig|1411691.4.peg.1940; -.
DR   EchoBASE; EB1302; -.
DR   eggNOG; COG0402; Bacteria.
DR   HOGENOM; CLU_031758_0_1_6; -.
DR   InParanoid; P25524; -.
DR   OMA; SDNCRDP; -.
DR   OrthoDB; 9815027at2; -.
DR   PhylomeDB; P25524; -.
DR   BioCyc; EcoCyc:CYTDEAM-MONOMER; -.
DR   BioCyc; MetaCyc:CYTDEAM-MONOMER; -.
DR   BRENDA; 3.5.4.1; 2026.
DR   EvolutionaryTrace; P25524; -.
DR   PRO; PR:P25524; -.
DR   Proteomes; UP000000318; Chromosome.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005829; C:cytosol; HDA:UniProtKB.
DR   GO; GO:0102480; F:5-fluorocytosine deaminase activity; IEA:UniProtKB-EC.
DR   GO; GO:0004131; F:cytosine deaminase activity; IDA:EcoCyc.
DR   GO; GO:0008198; F:ferrous iron binding; IDA:EcoCyc.
DR   GO; GO:0042802; F:identical protein binding; IDA:EcoCyc.
DR   GO; GO:0035888; F:isoguanine deaminase activity; IDA:EcoCyc.
DR   GO; GO:0008270; F:zinc ion binding; IDA:EcoCyc.
DR   GO; GO:0006209; P:cytosine catabolic process; IMP:EcoCyc.
DR   CDD; cd01293; Bact_CD; 1.
DR   Gene3D; 3.20.20.140; Metal-dependent hydrolases; 1.
DR   Gene3D; 2.30.40.10; Urease, subunit C, domain 1; 1.
DR   InterPro; IPR013108; Amidohydro_3.
DR   InterPro; IPR011059; Metal-dep_hydrolase_composite.
DR   InterPro; IPR032466; Metal_Hydrolase.
DR   PANTHER; PTHR32027; CYTOSINE DEAMINASE; 1.
DR   PANTHER; PTHR32027:SF0; CYTOSINE DEAMINASE; 1.
DR   Pfam; PF07969; Amidohydro_3; 1.
DR   SUPFAM; SSF51338; Composite domain of metallo-dependent hydrolases; 1.
DR   SUPFAM; SSF51556; Metallo-dependent hydrolases; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Cytosine metabolism; Direct protein sequencing; Hydrolase;
KW   Iron; Metal-binding; Reference proteome; Zinc.
FT   INIT_MET        1
FT                   /note="Removed"
FT                   /evidence="ECO:0000269|PubMed:1640834"
FT   CHAIN           2..427
FT                   /note="Cytosine deaminase"
FT                   /id="PRO_0000090002"
FT   ACT_SITE        218
FT                   /note="Proton donor"
FT                   /evidence="ECO:0000305|PubMed:11812140,
FT                   ECO:0000305|PubMed:21545144, ECO:0000305|PubMed:21604715"
FT   BINDING         62
FT                   /ligand="Fe(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29033"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:11812140,
FT                   ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:19487291"
FT   BINDING         62
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:21545144,
FT                   ECO:0000269|PubMed:21604715, ECO:0000269|Ref.14"
FT   BINDING         64
FT                   /ligand="Fe(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29033"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:11812140,
FT                   ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:19487291"
FT   BINDING         64
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:21545144,
FT                   ECO:0000269|PubMed:21604715, ECO:0000269|Ref.14"
FT   BINDING         157
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000269|PubMed:11812140,
FT                   ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:21545144,
FT                   ECO:0000269|PubMed:21604715"
FT   BINDING         215
FT                   /ligand="Fe(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29033"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:11812140,
FT                   ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:19487291"
FT   BINDING         215
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:21545144,
FT                   ECO:0000269|PubMed:21604715, ECO:0000269|Ref.14"
FT   BINDING         314
FT                   /ligand="Fe(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29033"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:11812140,
FT                   ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:19487291"
FT   BINDING         314
FT                   /ligand="Zn(2+)"
FT                   /ligand_id="ChEBI:CHEBI:29105"
FT                   /ligand_note="catalytic"
FT                   /evidence="ECO:0000269|PubMed:21545144,
FT                   ECO:0000269|PubMed:21604715, ECO:0000269|Ref.14"
FT   BINDING         320
FT                   /ligand="substrate"
FT                   /evidence="ECO:0000269|PubMed:11812140,
FT                   ECO:0000269|PubMed:15381761, ECO:0000269|PubMed:21545144"
FT   SITE            247
FT                   /note="Activates the nucleophilic water"
FT                   /evidence="ECO:0000305|PubMed:21545144,
FT                   ECO:0000305|PubMed:21604715"
FT   VARIANT         13
FT                   /note="R -> W (in strain: SO5076)"
FT   MUTAGEN         157
FT                   /note="Q->A,N: Less than 0.01% of wild-type enzymatic
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:21545144"
FT   MUTAGEN         218
FT                   /note="E->A,Q: Less than 0.01% of wild-type enzymatic
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:21545144"
FT   MUTAGEN         247
FT                   /note="H->A,N: Less than 0.01% of wild-type enzymatic
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:21545144"
FT   MUTAGEN         247
FT                   /note="H->Q: 200-fold decrease in catalytic efficiency."
FT                   /evidence="ECO:0000269|PubMed:21545144"
FT   MUTAGEN         314
FT                   /note="D->A: 17-fold decrease in catalytic efficiency with
FT                   cytosine as substrate and 2-fold increase in that with 5FC
FT                   as substrate. Shows increased sensitivity to 5FC."
FT                   /evidence="ECO:0000269|PubMed:15381761"
FT   MUTAGEN         314
FT                   /note="D->A: Less than 0.01% of wild-type enzymatic
FT                   activity."
FT                   /evidence="ECO:0000269|PubMed:21545144"
FT   MUTAGEN         314
FT                   /note="D->G,S: Still active towards cytosine. Shows
FT                   increased sensitivity to 5FC."
FT                   /evidence="ECO:0000269|PubMed:15381761"
FT   MUTAGEN         314
FT                   /note="D->K,R,H: Loss of enzymatic activity."
FT                   /evidence="ECO:0000269|PubMed:15381761"
FT   MUTAGEN         314
FT                   /note="D->N: 35000-fold decrease in catalytic efficiency."
FT                   /evidence="ECO:0000269|PubMed:21545144"
FT   STRAND          8..13
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          19..26
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          29..38
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          46..48
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          53..56
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          58..63
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   TURN            65..69
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          73..75
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           82..93
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           98..114
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          117..125
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           132..144
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   TURN            145..147
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          149..155
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          160..163
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           166..175
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          179..181
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           185..187
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          188..190
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           191..208
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          211..216
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           226..237
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           240..242
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          243..247
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           249..253
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           256..269
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          272..275
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           277..283
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   TURN            284..287
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           299..304
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          309..311
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          316..318
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           328..338
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           344..348
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           349..354
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           356..361
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          368..370
FT                   /evidence="ECO:0007829|PDB:3R0D"
FT   STRAND          373..375
FT                   /evidence="ECO:0007829|PDB:3RN6"
FT   STRAND          377..384
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   HELIX           385..391
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          396..400
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          403..407
FT                   /evidence="ECO:0007829|PDB:1RA0"
FT   STRAND          413..423
FT                   /evidence="ECO:0007829|PDB:1RA0"
SQ   SEQUENCE   427 AA;  47591 MW;  9F91A2C46B3B1E42 CRC64;
     MSNNALQTII NARLPGEEGL WQIHLQDGKI SAIDAQSGVM PITENSLDAE QGLVIPPFVE
     PHIHLDTTQT AGQPNWNQSG TLFEGIERWA ERKALLTHDD VKQRAWQTLK WQIANGIQHV
     RTHVDVSDAT LTALKAMLEV KQEVAPWIDL QIVAFPQEGI LSYPNGEALL EEALRLGADV
     VGAIPHFEFT REYGVESLHK TFALAQKYDR LIDVHCDEID DEQSRFVETV AALAHHEGMG
     ARVTASHTTA MHSYNGAYTS RLFRLLKMSG INFVANPLVN IHLQGRFDTY PKRRGITRVK
     EMLESGINVC FGHDDVFDPW YPLGTANMLQ VLHMGLHVCQ LMGYGQINDG LNLITHHSAR
     TLNLQDYGIA AGNSANLIIL PAENGFDALR RQVPVRYSVR GGKVIASTQP AQTTVYLEQP
     EAIDYKR
//