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 12-OCT-2022, entry version 79. 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}. 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; 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; 69756039; -. 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 //