PYRG_LACLM
ID PYRG_LACLM Reviewed; 535 AA.
AC O87761; A2RIH6;
DT 19-OCT-2002, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-2000, sequence version 2.
DT 03-AUG-2022, entry version 115.
DE RecName: Full=CTP synthase {ECO:0000255|HAMAP-Rule:MF_01227, ECO:0000303|PubMed:11500486};
DE EC=6.3.4.2 {ECO:0000255|HAMAP-Rule:MF_01227, ECO:0000269|PubMed:11500486};
DE AltName: Full=Cytidine 5'-triphosphate synthase {ECO:0000255|HAMAP-Rule:MF_01227};
DE AltName: Full=Cytidine triphosphate synthetase {ECO:0000255|HAMAP-Rule:MF_01227};
DE Short=CTP synthetase {ECO:0000255|HAMAP-Rule:MF_01227};
DE Short=CTPS {ECO:0000255|HAMAP-Rule:MF_01227};
DE AltName: Full=UTP--ammonia ligase {ECO:0000255|HAMAP-Rule:MF_01227};
GN Name=pyrG {ECO:0000255|HAMAP-Rule:MF_01227}; OrderedLocusNames=llmg_0467;
OS Lactococcus lactis subsp. cremoris (strain MG1363).
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Streptococcaceae;
OC Lactococcus; Lactococcus cremoris subsp. cremoris.
OX NCBI_TaxID=416870;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY REGULATION, DISRUPTION PHENOTYPE,
RP SUBUNIT, AND PATHWAY.
RC STRAIN=MG1363;
RX PubMed=11500486; DOI=10.1074/jbc.m100531200;
RA Wadskov-Hansen S.L.L., Willemoes M., Martinussen J., Hammer K., Neuhard J.,
RA Larsen S.;
RT "Cloning and verification of the Lactococcus lactis pyrG gene and
RT characterization of the gene product, CTP synthase.";
RL J. Biol. Chem. 276:38002-38009(2001).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=MG1363;
RX PubMed=17307855; DOI=10.1128/jb.01768-06;
RA Wegmann U., O'Connell-Motherway M., Zomer A., Buist G., Shearman C.,
RA Canchaya C., Ventura M., Goesmann A., Gasson M.J., Kuipers O.P.,
RA van Sinderen D., Kok J.;
RT "The complete genome sequence of the lactic acid bacterial paradigm
RT Lactococcus lactis subsp. cremoris MG1363.";
RL J. Bacteriol. 189:3256-3270(2007).
CC -!- FUNCTION: Catalyzes the ATP-dependent amination of UTP to CTP with
CC either L-glutamine or ammonia as the source of nitrogen. Is essential
CC for the synthesis of CTP de novo. Contrary to other bacterial CTP
CC synthases, the lactococcal enzyme is also able to convert dUTP to dCTP,
CC but this reaction may not play a significant physiological role
CC (PubMed:11500486). Regulates intracellular CTP levels through
CC interactions with the four ribonucleotide triphosphates (By
CC similarity). {ECO:0000255|HAMAP-Rule:MF_01227,
CC ECO:0000269|PubMed:11500486}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + H2O + L-glutamine + UTP = ADP + CTP + 2 H(+) + L-
CC glutamate + phosphate; Xref=Rhea:RHEA:26426, ChEBI:CHEBI:15377,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:29985, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:37563, ChEBI:CHEBI:43474, ChEBI:CHEBI:46398,
CC ChEBI:CHEBI:58359, ChEBI:CHEBI:456216; EC=6.3.4.2;
CC Evidence={ECO:0000255|HAMAP-Rule:MF_01227,
CC ECO:0000269|PubMed:11500486};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=H2O + L-glutamine = L-glutamate + NH4(+);
CC Xref=Rhea:RHEA:15889, ChEBI:CHEBI:15377, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:29985, ChEBI:CHEBI:58359; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_01227};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=ATP + NH4(+) + UTP = ADP + CTP + 2 H(+) + phosphate;
CC Xref=Rhea:RHEA:16597, ChEBI:CHEBI:15378, ChEBI:CHEBI:28938,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:37563, ChEBI:CHEBI:43474,
CC ChEBI:CHEBI:46398, ChEBI:CHEBI:456216; Evidence={ECO:0000255|HAMAP-
CC Rule:MF_01227, ECO:0000269|PubMed:11500486};
CC -!- ACTIVITY REGULATION: Allosterically activated by GTP, when glutamine is
CC the substrate (PubMed:11500486). GTP has no effect on the reaction when
CC ammonia is the substrate. The allosteric effector GTP functions by
CC stabilizing the protein conformation that binds the tetrahedral
CC intermediate(s) formed during glutamine hydrolysis (By similarity).
CC Also activated by magnesium (PubMed:11500486). Allosterically inhibited
CC by CTP (PubMed:11500486). {ECO:0000255|HAMAP-Rule:MF_01227,
CC ECO:0000269|PubMed:11500486}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.52 mM for L-glutamine {ECO:0000269|PubMed:11500486};
CC KM=42.9 mM for ammonia {ECO:0000269|PubMed:11500486};
CC -!- PATHWAY: Pyrimidine metabolism; CTP biosynthesis via de novo pathway;
CC CTP from UDP: step 2/2. {ECO:0000255|HAMAP-Rule:MF_01227,
CC ECO:0000269|PubMed:11500486}.
CC -!- SUBUNIT: Homotetramer. In contrast to E.coli CTP synthase, remains a
CC tetramer at dilute enzyme concentrations even in the absence of Mg(2+),
CC ATP and UTP. {ECO:0000269|PubMed:11500486}.
CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene require cytidine for
CC growth, proving that in L.lactis, the pyrG product is the only enzyme
CC responsible for the amination of UTP to CTP.
CC {ECO:0000269|PubMed:11500486}.
CC -!- MISCELLANEOUS: CTPSs have evolved a hybrid strategy for distinguishing
CC between UTP and CTP. The overlapping regions of the product feedback
CC inhibitory and substrate sites recognize a common feature in both
CC compounds, the triphosphate moiety. To differentiate isosteric
CC substrate and product pyrimidine rings, an additional pocket far from
CC the expected kinase/ligase catalytic site, specifically recognizes the
CC cytosine and ribose portions of the product inhibitor.
CC {ECO:0000255|HAMAP-Rule:MF_01227}.
CC -!- SIMILARITY: Belongs to the CTP synthase family. {ECO:0000255|HAMAP-
CC Rule:MF_01227}.
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DR EMBL; AJ010153; CAA09021.2; -; Genomic_DNA.
DR EMBL; AM406671; CAL97071.1; -; Genomic_DNA.
DR RefSeq; WP_011834508.1; NZ_WJVF01000001.1.
DR AlphaFoldDB; O87761; -.
DR SMR; O87761; -.
DR STRING; 416870.llmg_0467; -.
DR MEROPS; C26.964; -.
DR EnsemblBacteria; CAL97071; CAL97071; llmg_0467.
DR KEGG; llm:llmg_0467; -.
DR eggNOG; COG0504; Bacteria.
DR HOGENOM; CLU_011675_5_0_9; -.
DR OMA; EFNNAYR; -.
DR PhylomeDB; O87761; -.
DR BioCyc; LLAC416870:LLMG_RS02375-MON; -.
DR SABIO-RK; O87761; -.
DR UniPathway; UPA00159; UER00277.
DR Proteomes; UP000000364; Chromosome.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0003883; F:CTP synthase activity; IEA:UniProtKB-UniRule.
DR GO; GO:0004359; F:glutaminase activity; IEA:RHEA.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0044210; P:'de novo' CTP biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0006541; P:glutamine metabolic process; IEA:UniProtKB-KW.
DR CDD; cd03113; CTPS_N; 1.
DR CDD; cd01746; GATase1_CTP_Synthase; 1.
DR Gene3D; 3.40.50.300; -; 1.
DR Gene3D; 3.40.50.880; -; 1.
DR HAMAP; MF_01227; PyrG; 1.
DR InterPro; IPR029062; Class_I_gatase-like.
DR InterPro; IPR004468; CTP_synthase.
DR InterPro; IPR017456; CTP_synthase_N.
DR InterPro; IPR017926; GATASE.
DR InterPro; IPR033828; GATase1_CTP_Synthase.
DR InterPro; IPR027417; P-loop_NTPase.
DR PANTHER; PTHR11550; PTHR11550; 1.
DR Pfam; PF06418; CTP_synth_N; 1.
DR Pfam; PF00117; GATase; 1.
DR SUPFAM; SSF52317; SSF52317; 1.
DR SUPFAM; SSF52540; SSF52540; 1.
DR TIGRFAMs; TIGR00337; PyrG; 1.
DR PROSITE; PS51273; GATASE_TYPE_1; 1.
PE 1: Evidence at protein level;
KW ATP-binding; Glutamine amidotransferase; Ligase; Magnesium; Metal-binding;
KW Nucleotide-binding; Pyrimidine biosynthesis.
FT CHAIN 1..535
FT /note="CTP synthase"
FT /id="PRO_0000138192"
FT DOMAIN 293..535
FT /note="Glutamine amidotransferase type-1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT REGION 1..268
FT /note="Amidoligase domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT ACT_SITE 382
FT /note="Nucleophile; for glutamine hydrolysis"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT ACT_SITE 509
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT ACT_SITE 511
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 14
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 14
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 15..20
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 55
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 72
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 72
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 142
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 149..151
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 189..194
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 189..194
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 225
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 225
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 243
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 355
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 383..386
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 406
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 464
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
SQ SEQUENCE 535 AA; 59455 MW; 045724E0FEDE5B4E CRC64;
MSTKYIFVTG GGTSSMGKGI VAASLGRLLK NRGLKVTVQK FDPYLNIDPG TMSPYQHGEV
FVTDDGAETD LDLGHYERFI DINLNKYSNV TSGKVYSEIL RKERKGEYLG ATVQMVPHVT
NMLKEKIKRA ATTTDADIII TEVGGTVGDM ESLPFIEALR QMKAEVGADN VMYIHTVPIL
HLRAAGELKT KIAQNATKTL REYGIQANML VLRSEVPITT EMRDKIAMFC DVAPEAVIQS
LDVEHLYQIP LNLQAQNMDQ IVCDHLKLDA PKADMAEWSA MVDHVMNLKK KVKIALVGKY
VELPDAYISV TEALKHAGYA SDAEVDINWV NANDVTDENV AELVGDAAGI IVPGGFGQRG
TEGKIAAIKY ARENDVPMLG ICLGMQLTAV EFARNVLGLE GAHSFELDPE TKYPVIDIMR
DQVDVEDMGG TLRLGLYPAK LKNGSRAKAA YNDAEVVQRR HRHRYEFNNK YREDFEKAGF
VFSGVSPDNR LVEIVELSGK KFFVACQYHP ELQSRPNRPE ELYTEFIRVA VENSK
