CAH_AZOC5
ID CAH_AZOC5 Reviewed; 356 AA.
AC A8IKD2;
DT 10-MAY-2017, integrated into UniProtKB/Swiss-Prot.
DT 04-DEC-2007, sequence version 1.
DT 03-AUG-2022, entry version 67.
DE RecName: Full=Cyanuric acid amidohydrolase {ECO:0000255|HAMAP-Rule:MF_01989};
DE Short=CAH {ECO:0000255|HAMAP-Rule:MF_01989};
DE EC=3.5.2.15 {ECO:0000255|HAMAP-Rule:MF_01989, ECO:0000269|PubMed:22730121};
GN OrderedLocusNames=AZC_3892;
OS Azorhizobium caulinodans (strain ATCC 43989 / DSM 5975 / JCM 20966 / LMG
OS 6465 / NBRC 14845 / NCIMB 13405 / ORS 571).
OC Bacteria; Proteobacteria; Alphaproteobacteria; Hyphomicrobiales;
OC Xanthobacteraceae; Azorhizobium.
OX NCBI_TaxID=438753;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 43989 / DSM 5975 / JCM 20966 / LMG 6465 / NBRC 14845 / NCIMB
RC 13405 / ORS 571;
RA Lee K.B., Backer P.D., Aono T., Liu C.T., Suzuki S., Suzuki T., Kaneko T.,
RA Yamada M., Tabata S., Kupfer D.M., Najar F.Z., Wiley G.B., Roe B.,
RA Binnewies T., Ussery D., Vereecke D., Gevers D., Holsters M., Oyaizu H.;
RT "Complete genome sequence of the nitrogen-fixing bacterium Azorhizobium
RT caulinodans ORS571.";
RL Submitted (APR-2007) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, AND BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=22730121; DOI=10.1128/jb.00791-12;
RA Seffernick J.L., Erickson J.S., Cameron S.M., Cho S., Dodge A.G.,
RA Richman J.E., Sadowsky M.J., Wackett L.P.;
RT "Defining sequence space and reaction products within the cyanuric acid
RT hydrolase (AtzD)/barbiturase protein family.";
RL J. Bacteriol. 194:4579-4588(2012).
RN [3]
RP SUBUNIT.
RX PubMed=23908033; DOI=10.1107/s1744309113017077;
RA Cho S., Shi K., Wackett L.P., Aihara H.;
RT "Crystallization and preliminary X-ray diffraction studies of cyanuric acid
RT hydrolase from Azorhizobium caulinodans.";
RL Acta Crystallogr. F 69:880-883(2013).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.70 ANGSTROMS) IN COMPLEX WITH MAGNESIUM AND
RP INHIBITOR, ACTIVE SITE, AND MUTAGENESIS OF LYS-40; SER-79; LYS-156;
RP SER-226; LYS-285 AND SER-333.
RX PubMed=24915109; DOI=10.1371/journal.pone.0099349;
RA Cho S., Shi K., Seffernick J.L., Dodge A.G., Wackett L.P., Aihara H.;
RT "Cyanuric acid hydrolase from Azorhizobium caulinodans ORS 571: crystal
RT structure and insights into a new class of Ser-Lys dyad proteins.";
RL PLoS ONE 9:E99349-E99349(2014).
CC -!- FUNCTION: Responsible for the hydrolysis of cyanuric acid, an
CC intermediate formed during catabolism of s-triazine based compounds in
CC herbicides such as atrazine and polymers such as melamine. Catalyzes
CC the hydrolytic opening of the s-triazine ring of cyanuric acid (2,4,6-
CC trihydroxy-s-triazine) to yield carbon dioxide and carboxybiuret, which
CC spontaneously decarboxylates to biuret. {ECO:0000255|HAMAP-
CC Rule:MF_01989, ECO:0000269|PubMed:22730121}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=cyanurate + H2O = 1-carboxybiuret + H(+);
CC Xref=Rhea:RHEA:70363, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:38028, ChEBI:CHEBI:142864; EC=3.5.2.15;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01989,
CC ECO:0000269|PubMed:22730121};
CC -!- ACTIVITY REGULATION: Inhibited by barbituric acid.
CC {ECO:0000250|UniProtKB:P58329, ECO:0000255|HAMAP-Rule:MF_01989}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=370 uM for cyanuric acid {ECO:0000269|PubMed:22730121};
CC Note=kcat is 50 sec(-1) with cyanuric acid as substrate.
CC {ECO:0000269|PubMed:22730121};
CC -!- PATHWAY: Xenobiotic degradation; atrazine degradation; biuret from
CC cyanurate: step 1/1. {ECO:0000255|HAMAP-Rule:MF_01989}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000255|HAMAP-Rule:MF_01989,
CC ECO:0000269|PubMed:23908033}.
CC -!- DOMAIN: The monomer structure is formed from three repeating units
CC (RUs) that share the same structure as one another. The monomer, the
CC active site and substrate all possess threefold rotational symmetry, to
CC the extent that the active site possesses three potential Ser-Lys
CC catalytic dyads. It is possible that any or all of the three active-
CC site serines may act as nucleophile (albeit only one can do so per
CC catalytic cycle). Mutant, bioinformatic and structural data propose one
CC specific dyad being involved in catalysis. {ECO:0000255|HAMAP-
CC Rule:MF_01989, ECO:0000305|PubMed:24915109}.
CC -!- SIMILARITY: Belongs to the cyclic amide hydrolase (CyAH) family.
CC {ECO:0000255|HAMAP-Rule:MF_01989, ECO:0000305}.
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DR EMBL; AP009384; BAF89890.1; -; Genomic_DNA.
DR RefSeq; WP_012172412.1; NC_009937.1.
DR PDB; 4NQ3; X-ray; 2.70 A; A/B=1-356.
DR PDBsum; 4NQ3; -.
DR AlphaFoldDB; A8IKD2; -.
DR SMR; A8IKD2; -.
DR STRING; 438753.AZC_3892; -.
DR EnsemblBacteria; BAF89890; BAF89890; AZC_3892.
DR KEGG; azc:AZC_3892; -.
DR eggNOG; ENOG502Z8BS; Bacteria.
DR HOGENOM; CLU_808206_0_0_5; -.
DR OMA; GRYRIGH; -.
DR OrthoDB; 984718at2; -.
DR UniPathway; UPA00008; UER00502.
DR Proteomes; UP000000270; Chromosome.
DR GO; GO:0018753; F:cyanuric acid amidohydrolase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0019381; P:atrazine catabolic process; IEA:UniProtKB-UniRule.
DR Gene3D; 3.30.1330.160; -; 1.
DR Gene3D; 3.30.1330.170; -; 1.
DR Gene3D; 3.30.1330.180; -; 1.
DR HAMAP; MF_01989; Cyc_amidohydrol; 1.
DR InterPro; IPR014086; AtzD/Barbiturase.
DR InterPro; IPR043008; AtzD/Barbiturase_RUA.
DR InterPro; IPR043006; AtzD/Barbiturase_RUB.
DR InterPro; IPR043007; AtzD/Barbiturase_RUC.
DR Pfam; PF09663; Amido_AtzD_TrzD; 1.
DR TIGRFAMs; TIGR02714; amido_AtzD_TrzD; 1.
PE 1: Evidence at protein level;
KW 3D-structure; Hydrolase; Magnesium; Metal-binding; Reference proteome.
FT CHAIN 1..356
FT /note="Cyanuric acid amidohydrolase"
FT /id="PRO_0000439909"
FT REGION 1..99
FT /note="RU A"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989"
FT REGION 106..243
FT /note="RU B"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989"
FT REGION 249..356
FT /note="RU C"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989"
FT ACT_SITE 156
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT ACT_SITE 226
FT /note="Nucleophile"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 52
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 79..80
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 188
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 226..227
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 287
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 314
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 333..334
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 336
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 339
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 340
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 341
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT BINDING 344
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989,
FT ECO:0000269|PubMed:24915109"
FT SITE 310
FT /note="Important for substrate specificity"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01989"
FT MUTAGEN 40
FT /note="K->A: Reduces catalytic activity 1e4-fold."
FT /evidence="ECO:0000269|PubMed:24915109"
FT MUTAGEN 79
FT /note="S->A: Reduces catalytic activity 1e8-fold."
FT /evidence="ECO:0000269|PubMed:24915109"
FT MUTAGEN 156
FT /note="K->A: Reduces catalytic activity 1e4-fold."
FT /evidence="ECO:0000269|PubMed:24915109"
FT MUTAGEN 226
FT /note="S->A: Reduces catalytic activity 1e9-fold."
FT /evidence="ECO:0000269|PubMed:24915109"
FT MUTAGEN 285
FT /note="K->A: Reduces catalytic activity 1e4-fold."
FT /evidence="ECO:0000269|PubMed:24915109"
FT MUTAGEN 333
FT /note="S->A: Reduces catalytic activity 1e8-fold."
FT /evidence="ECO:0000269|PubMed:24915109"
FT STRAND 2..10
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 18..25
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 31..33
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 34..43
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 51..67
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 68..73
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 74..80
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 89..97
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 106..113
FT /evidence="ECO:0007829|PDB:4NQ3"
FT TURN 119..123
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 125..141
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 147..149
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 150..157
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 162..170
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 180..199
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 204..206
FT /evidence="ECO:0007829|PDB:4NQ3"
FT TURN 209..214
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 220..227
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 235..242
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 246..258
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 263..271
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 281..286
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 292..294
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 305..307
FT /evidence="ECO:0007829|PDB:4NQ3"
FT HELIX 309..325
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 331..334
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 339..341
FT /evidence="ECO:0007829|PDB:4NQ3"
FT STRAND 345..352
FT /evidence="ECO:0007829|PDB:4NQ3"
SQ SEQUENCE 356 AA; 36040 MW; 231B48FE327A4E3C CRC64;
MPIAKVHRIA TASPDDVSGL AAAIATGAIA PAGILAIFGK TEGNGCVNDF SRGFAVQSLQ
MLLRGHMGAA ADEVCLVMSG GTEGGMSPHF LVFERAEGNA PEAAPALAIG RAHTPDLPFE
ALGRMGQVRM VAQAVRRAMA AAGITDPEDV HFVQVKCPLL TAMRVKEAEA RGATTATSDT
LKSMGLSRGA SALGIALALG EVAEDALSDA VICADYGLWS ARASCSSGIE LLGHEIVVLG
MSEGWSGPLA IAHGVMADAI DVTPVKAALS ALGAEAGEAT IVLAKAEPSR SGRIRGKRHT
MLDDSDISPT RHARAFVAGA LAGVVGHTEI YVSGGGEHQG PDGGGPVAVI AARTMG
