PYRG_THET8
ID PYRG_THET8 Reviewed; 550 AA.
AC Q5SIA8;
DT 12-DEC-2006, integrated into UniProtKB/Swiss-Prot.
DT 21-DEC-2004, sequence version 1.
DT 03-AUG-2022, entry version 108.
DE RecName: Full=CTP synthase {ECO:0000255|HAMAP-Rule:MF_01227};
DE EC=6.3.4.2 {ECO:0000255|HAMAP-Rule:MF_01227, ECO:0000269|PubMed:15296735};
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, ECO:0000303|PubMed:15296735};
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=TTHA1466;
OS Thermus thermophilus (strain ATCC 27634 / DSM 579 / HB8).
OC Bacteria; Deinococcus-Thermus; Deinococci; Thermales; Thermaceae; Thermus.
OX NCBI_TaxID=300852;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 27634 / DSM 579 / HB8;
RA Masui R., Kurokawa K., Nakagawa N., Tokunaga F., Koyama Y., Shibata T.,
RA Oshima T., Yokoyama S., Yasunaga T., Kuramitsu S.;
RT "Complete genome sequence of Thermus thermophilus HB8.";
RL Submitted (NOV-2004) to the EMBL/GenBank/DDBJ databases.
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.15 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH
RP L-GLUTAMINE, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES,
RP AND SUBUNIT.
RC STRAIN=ATCC 27634 / DSM 579 / HB8;
RX PubMed=15296735; DOI=10.1016/j.str.2004.05.013;
RA Goto M., Omi R., Nakagawa N., Miyahara I., Hirotsu K.;
RT "Crystal structures of CTP synthetase reveal ATP, UTP, and glutamine
RT binding sites.";
RL Structure 12:1413-1423(2004).
CC -!- FUNCTION: Catalyzes the ATP-dependent amination of UTP to CTP with
CC either L-glutamine or ammonia as the source of nitrogen
CC (PubMed:15296735). 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:15296735}.
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:15296735};
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, ECO:0000305|PubMed:15296735};
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:0000305|PubMed:15296735};
CC -!- ACTIVITY REGULATION: Allosterically activated by GTP, when glutamine is
CC the substrate (PubMed:15296735). 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. Inhibited by the
CC product CTP, via allosteric rather than competitive inhibition (By
CC similarity). {ECO:0000255|HAMAP-Rule:MF_01227,
CC ECO:0000269|PubMed:15296735}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=0.24 mM for L-glutamine (at unsaturating levels of ATP and UTP
CC (0.5 mM)) {ECO:0000269|PubMed:15296735};
CC KM=0.22 mM for L-glutamine (in the presence of GTP)
CC {ECO:0000269|PubMed:15296735};
CC KM=0.37 mM for UTP {ECO:0000269|PubMed:15296735};
CC Note=kcat is 7.4 sec(-1) toward L-glutamine at unsaturating levels of
CC ATP and UTP (0.5 mM) and 40 sec(-1) in the presence of GTP. kcat is
CC 260 sec(-1) toward UTP. {ECO:0000269|PubMed:15296735};
CC -!- PATHWAY: Pyrimidine metabolism; CTP biosynthesis via de novo pathway;
CC CTP from UDP: step 2/2. {ECO:0000255|HAMAP-Rule:MF_01227}.
CC -!- SUBUNIT: Homotetramer in the presence of UTP and ATP. Is in a protein
CC concentration-dependent equilibrium between monomer, dimer, and
CC tetramer in the absence of UTP and ATP. {ECO:0000269|PubMed:15296735}.
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; AP008226; BAD71289.1; -; Genomic_DNA.
DR RefSeq; WP_011228700.1; NC_006461.1.
DR RefSeq; YP_144732.1; NC_006461.1.
DR PDB; 1VCM; X-ray; 2.35 A; A=1-550.
DR PDB; 1VCN; X-ray; 2.25 A; A=1-550.
DR PDB; 1VCO; X-ray; 2.15 A; A=1-550.
DR PDBsum; 1VCM; -.
DR PDBsum; 1VCN; -.
DR PDBsum; 1VCO; -.
DR AlphaFoldDB; Q5SIA8; -.
DR SMR; Q5SIA8; -.
DR STRING; 300852.55772848; -.
DR MEROPS; C26.964; -.
DR EnsemblBacteria; BAD71289; BAD71289; BAD71289.
DR GeneID; 3169502; -.
DR KEGG; ttj:TTHA1466; -.
DR PATRIC; fig|300852.9.peg.1440; -.
DR eggNOG; COG0504; Bacteria.
DR HOGENOM; CLU_011675_5_0_0; -.
DR OMA; EFNNAYR; -.
DR PhylomeDB; Q5SIA8; -.
DR BRENDA; 6.3.4.2; 7852.
DR UniPathway; UPA00159; UER00277.
DR EvolutionaryTrace; Q5SIA8; -.
DR Proteomes; UP000000532; 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 3D-structure; ATP-binding; Glutamine amidotransferase; Ligase; Magnesium;
KW Metal-binding; Nucleotide-binding; Pyrimidine biosynthesis;
KW Reference proteome.
FT CHAIN 1..550
FT /note="CTP synthase"
FT /id="PRO_0000266251"
FT DOMAIN 302..549
FT /note="Glutamine amidotransferase type-1"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT REGION 1..277
FT /note="Amidoligase domain"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT ACT_SITE 391
FT /note="Nucleophile; for glutamine hydrolysis"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227,
FT ECO:0000305|PubMed:15296735"
FT ACT_SITE 522
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227,
FT ECO:0000305|PubMed:15296735"
FT ACT_SITE 524
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227,
FT ECO:0000305|PubMed:15296735"
FT BINDING 23
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 23
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 24..29
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 64
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000269|PubMed:15296735,
FT ECO:0007744|PDB:1VCO"
FT BINDING 81
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 81
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 151
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 158..160
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 198..203
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 198..203
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 234
FT /ligand="CTP"
FT /ligand_id="ChEBI:CHEBI:37563"
FT /ligand_note="allosteric inhibitor"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 234
FT /ligand="UTP"
FT /ligand_id="ChEBI:CHEBI:46398"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 252
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01227"
FT BINDING 364
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000269|PubMed:15296735,
FT ECO:0007744|PDB:1VCO"
FT BINDING 392..395
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000269|PubMed:15296735,
FT ECO:0007744|PDB:1VCO"
FT BINDING 415
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000269|PubMed:15296735,
FT ECO:0007744|PDB:1VCO"
FT BINDING 472
FT /ligand="L-glutamine"
FT /ligand_id="ChEBI:CHEBI:58359"
FT /evidence="ECO:0000269|PubMed:15296735,
FT ECO:0007744|PDB:1VCO"
FT STRAND 13..19
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 21..23
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 27..39
FT /evidence="ECO:0007829|PDB:1VCO"
FT TURN 40..42
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 45..51
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 58..60
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 81..89
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 95..97
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 98..100
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 101..113
FT /evidence="ECO:0007829|PDB:1VCO"
FT TURN 114..119
FT /evidence="ECO:0007829|PDB:1VCO"
FT TURN 124..126
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 127..142
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 146..152
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 159..161
FT /evidence="ECO:0007829|PDB:1VCN"
FT HELIX 162..169
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 171..175
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 179..187
FT /evidence="ECO:0007829|PDB:1VCO"
FT TURN 192..195
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 200..211
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 217..225
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 229..239
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 243..245
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 246..250
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 257..265
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 267..274
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 285..295
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 298..308
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 314..316
FT /evidence="ECO:0007829|PDB:1VCN"
FT HELIX 317..329
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 332..340
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 341..343
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 349..352
FT /evidence="ECO:0007829|PDB:1VCO"
FT TURN 353..355
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 359..361
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 370..382
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 387..390
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 392..404
FT /evidence="ECO:0007829|PDB:1VCO"
FT TURN 414..416
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 423..428
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 430..432
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 442..450
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 455..460
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 463..475
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 477..486
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 488..493
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 497..501
FT /evidence="ECO:0007829|PDB:1VCM"
FT STRAND 505..510
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 513..521
FT /evidence="ECO:0007829|PDB:1VCO"
FT HELIX 523..526
FT /evidence="ECO:0007829|PDB:1VCO"
FT STRAND 529..531
FT /evidence="ECO:0007829|PDB:1VCM"
FT HELIX 534..546
FT /evidence="ECO:0007829|PDB:1VCO"
SQ SEQUENCE 550 AA; 61003 MW; F3D20D9FC50DFDF8 CRC64;
MNGSADAGPR PRKYVFITGG VVSSLGKGIL TSSLGALLRA RGYRVTAIKI DPYVNVDAGT
MRPYEHGEVF VTADGAETDL DIGHYERFLD MDLSRGNNLT TGQVYLSVIQ KERRGEYLSQ
TVQVIPHITD EIKERIRKVA EEQKAEIVVV EVGGTVGDIE SLPFLEAIRQ FRFDEGEGNT
LYLHLTLVPY LETSEEFKTK PTQHSVATLR GVGIQPDILV LRSARPVPEE VRRKVALFTN
VRPGHVFSSP TVEHLYEVPL LLEEQGLGRA VERALGLEAV IPNLSFWQEA VRVLKHPERT
VKIAIAGKYV KMPDAYLSLL EALRHAGIKN RARVEVKWVD AESLEAADLD EAFRDVSGIL
VPGGFGVRGI EGKVRAAQYA RERKIPYLGI CLGLQIAVIE FARNVAGLKG ANSTEFDPHT
PHPVIDLMPE QLEVEGLGGT MRLGDWPMRI KPGTLLHRLY GKEEVLERHR HRYEVNPLYV
DGLERAGLVV SATTPGMRGR GAGLVEAIEL KDHPFFLGLQ SHPEFKSRPM RPSPPFVGFV
EAALAYQERA
