PYRC_KLEP7
ID PYRC_KLEP7 Reviewed; 348 AA.
AC A6T7D6;
DT 15-JAN-2008, integrated into UniProtKB/Swiss-Prot.
DT 21-AUG-2007, sequence version 1.
DT 03-AUG-2022, entry version 78.
DE RecName: Full=Dihydroorotase {ECO:0000255|HAMAP-Rule:MF_00219};
DE Short=DHOase {ECO:0000255|HAMAP-Rule:MF_00219};
DE EC=3.5.2.3 {ECO:0000255|HAMAP-Rule:MF_00219, ECO:0000269|PubMed:20676924};
GN Name=pyrC {ECO:0000255|HAMAP-Rule:MF_00219};
GN OrderedLocusNames=KPN78578_10460; ORFNames=KPN_01074;
OS Klebsiella pneumoniae subsp. pneumoniae (strain ATCC 700721 / MGH 78578).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Klebsiella/Raoultella group; Klebsiella.
OX NCBI_TaxID=272620;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 700721 / MGH 78578;
RG The Klebsiella pneumonia Genome Sequencing Project;
RA McClelland M., Sanderson E.K., Spieth J., Clifton W.S., Latreille P.,
RA Sabo A., Pepin K., Bhonagiri V., Porwollik S., Ali J., Wilson R.K.;
RL Submitted (SEP-2006) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES,
RP SUBUNIT, AND MUTAGENESIS OF HIS-17; HIS-19; LYS-103; HIS-140; HIS-178 AND
RP ASP-251.
RC STRAIN=ATCC 13883 / DSM 30104 / JCM 1662 / NBRC 14940 / NCIMB 13281 / NCTC
RC 9633;
RX PubMed=20676924; DOI=10.1007/s10930-010-9272-2;
RA Wang C.C., Tsau H.W., Chen W.T., Huang C.Y.;
RT "Identification and characterization of a putative dihydroorotase,
RT KPN01074, from Klebsiella pneumoniae.";
RL Protein J. 29:445-452(2010).
CC -!- FUNCTION: Catalyzes the reversible cyclization of carbamoyl aspartate
CC to dihydroorotate. {ECO:0000255|HAMAP-Rule:MF_00219,
CC ECO:0000269|PubMed:20676924}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(S)-dihydroorotate + H2O = H(+) + N-carbamoyl-L-aspartate;
CC Xref=Rhea:RHEA:24296, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:30864, ChEBI:CHEBI:32814; EC=3.5.2.3;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00219,
CC ECO:0000269|PubMed:20676924};
CC -!- COFACTOR:
CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_00219};
CC Name=Co(2+); Xref=ChEBI:CHEBI:48828;
CC Evidence={ECO:0000269|PubMed:20676924};
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:20676924};
CC Name=Ni(2+); Xref=ChEBI:CHEBI:49786;
CC Evidence={ECO:0000269|PubMed:20676924};
CC Note=Binds 2 Zn(2+) ions per subunit (By similarity). In vitro, shows
CC higher activity with Co(2+), Mg(2+), Ni(2+) and Mn(2+) than with Zn(2+)
CC (PubMed:20676924). {ECO:0000255|HAMAP-Rule:MF_00219,
CC ECO:0000269|PubMed:20676924};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.04 mM for dihydroorotate {ECO:0000269|PubMed:20676924};
CC Vmax=8.87 umol/min/mg enzyme {ECO:0000269|PubMed:20676924};
CC pH dependence:
CC Optimum pH is 9.0. {ECO:0000269|PubMed:20676924};
CC Temperature dependence:
CC Optimum temperature is around 60 degrees Celsius.
CC {ECO:0000269|PubMed:20676924};
CC -!- PATHWAY: Pyrimidine metabolism; UMP biosynthesis via de novo pathway;
CC (S)-dihydroorotate from bicarbonate: step 3/3. {ECO:0000255|HAMAP-
CC Rule:MF_00219}.
CC -!- SUBUNIT: Homodimer. {ECO:0000255|HAMAP-Rule:MF_00219,
CC ECO:0000269|PubMed:20676924}.
CC -!- SIMILARITY: Belongs to the metallo-dependent hydrolases superfamily.
CC DHOase family. Class II DHOase subfamily. {ECO:0000255|HAMAP-
CC Rule:MF_00219, ECO:0000305}.
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DR EMBL; CP000647; ABR76507.1; -; Genomic_DNA.
DR RefSeq; WP_004176586.1; NC_009648.1.
DR AlphaFoldDB; A6T7D6; -.
DR SMR; A6T7D6; -.
DR STRING; 272620.KPN_01074; -.
DR MEROPS; M38.A02; -.
DR EnsemblBacteria; ABR76507; ABR76507; KPN_01074.
DR GeneID; 61332269; -.
DR KEGG; kpn:KPN_01074; -.
DR HOGENOM; CLU_041558_1_0_6; -.
DR OMA; TLHHISM; -.
DR BRENDA; 3.5.2.3; 2814.
DR UniPathway; UPA00070; UER00117.
DR Proteomes; UP000000265; Chromosome.
DR GO; GO:0004151; F:dihydroorotase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0008270; F:zinc ion binding; IEA:UniProtKB-UniRule.
DR GO; GO:0044205; P:'de novo' UMP biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0019856; P:pyrimidine nucleobase biosynthetic process; IEA:InterPro.
DR CDD; cd01294; DHOase; 1.
DR HAMAP; MF_00219; PyrC_classII; 1.
DR InterPro; IPR006680; Amidohydro-rel.
DR InterPro; IPR004721; DHOdimr.
DR InterPro; IPR002195; Dihydroorotase_CS.
DR InterPro; IPR032466; Metal_Hydrolase.
DR PANTHER; PTHR43137; PTHR43137; 1.
DR Pfam; PF01979; Amidohydro_1; 1.
DR PIRSF; PIRSF001237; DHOdimr; 1.
DR SUPFAM; SSF51556; SSF51556; 1.
DR TIGRFAMs; TIGR00856; pyrC_dimer; 1.
DR PROSITE; PS00483; DIHYDROOROTASE_2; 1.
PE 1: Evidence at protein level;
KW Hydrolase; Metal-binding; Pyrimidine biosynthesis; Reference proteome;
KW Zinc.
FT CHAIN 1..348
FT /note="Dihydroorotase"
FT /id="PRO_1000024020"
FT ACT_SITE 251
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 17
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 19..21
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 19
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 45
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 103
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /note="via carbamate group"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 103
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /note="via carbamate group"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 140
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 140
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 178
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="2"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 223
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 251
FT /ligand="Zn(2+)"
FT /ligand_id="ChEBI:CHEBI:29105"
FT /ligand_label="1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 255
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT BINDING 267
FT /ligand="substrate"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT MOD_RES 103
FT /note="N6-carboxylysine"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_00219"
FT MUTAGEN 17
FT /note="H->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:20676924"
FT MUTAGEN 19
FT /note="H->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:20676924"
FT MUTAGEN 103
FT /note="K->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:20676924"
FT MUTAGEN 140
FT /note="H->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:20676924"
FT MUTAGEN 178
FT /note="H->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:20676924"
FT MUTAGEN 251
FT /note="D->A: Lack of activity."
FT /evidence="ECO:0000269|PubMed:20676924"
FT MUTAGEN 251
FT /note="D->E: 4-fold increase in activity."
FT /evidence="ECO:0000269|PubMed:20676924"
SQ SEQUENCE 348 AA; 38821 MW; 2B841BB50B30506D CRC64;
MTAQSQVLKI RRPDDWHIHL RDDDMLKTVV PYTSEFYGRA IVMPNLVPPV TTVAAAIAYR
QRIMDAVPAG HDFTPLMTCY LTDSLDPAEL ERGFNEGVFT AAKLYPANAT TNSSHGVTST
DAIMPVLERM EKLGMPLLVH GEVTHAEIDI FDREARFIET VMEPLRQRLP GLKVVFEHIT
TKDAAEYVRD GNELLAATIT PQHLMFNRNH MLVGGIRPHL YCLPVLKRNI HQQALRELVA
SGFSRAFLGT DSAPHARHRK EASCGCAGCF NAPTALGSYA TVFEEMNALQ HFEAFCSLNG
PRFYGLPVNE SYVELVREET TVVDSIALPN DTLVPFLAGE TVRWTVKK
