CODA_ECOLI
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 03-AUG-2022, entry version 167.
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; Proteobacteria; 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; P25524; -.
DR PRIDE; P25524; -.
DR EnsemblBacteria; AAC73440; AAC73440; b0337.
DR EnsemblBacteria; BAE76119; BAE76119; BAE76119.
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; YGLPRVN; -.
DR PhylomeDB; P25524; -.
DR BioCyc; EcoCyc:CYTDEAM-MON; -.
DR BioCyc; MetaCyc:CYTDEAM-MON; -.
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:0016814; F:hydrolase activity, acting on carbon-nitrogen (but not peptide) bonds, in cyclic amidines; IBA:GO_Central.
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 Gene3D; 2.30.40.10; -; 1.
DR InterPro; IPR013108; Amidohydro_3.
DR InterPro; IPR011059; Metal-dep_hydrolase_composite.
DR InterPro; IPR032466; Metal_Hydrolase.
DR Pfam; PF07969; Amidohydro_3; 1.
DR SUPFAM; SSF51338; SSF51338; 1.
DR SUPFAM; SSF51556; SSF51556; 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
