ID DRTS_TRYCR Reviewed; 521 AA. AC Q27793; DT 01-NOV-1997, integrated into UniProtKB/Swiss-Prot. DT 01-MAY-1997, sequence version 2. DT 24-JAN-2024, entry version 114. DE RecName: Full=Bifunctional dihydrofolate reductase-thymidylate synthase; DE Short=DHFR-TS; DE Includes: DE RecName: Full=Dihydrofolate reductase; DE EC=1.5.1.3; DE Includes: DE RecName: Full=Thymidylate synthase; DE EC=2.1.1.45; OS Trypanosoma cruzi. OC Eukaryota; Discoba; Euglenozoa; Kinetoplastea; Metakinetoplastina; OC Trypanosomatida; Trypanosomatidae; Trypanosoma; Schizotrypanum. OX NCBI_TaxID=5693; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=Y; RX PubMed=7969266; DOI=10.1016/0166-6851(94)90076-0; RA Reche P., Arrebola R., Olmo A., Santi D.V., Gonzalez-Pacanowska D., RA Ruiz-Perez L.M.; RT "Cloning and expression of the dihydrofolate reductase-thymidylate synthase RT gene from Trypanosoma cruzi."; RL Mol. Biochem. Parasitol. 65:247-258(1994). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=Y; RX PubMed=8920005; DOI=10.1016/0166-6851(95)02557-x; RA Reche P., Arrebola R., Santi D.V., Gonzalez-Pacanowska D., Ruiz-Perez L.M.; RT "Expression and characterization of the Trypanosoma cruzi dihydrofolate RT reductase domain."; RL Mol. Biochem. Parasitol. 76:175-185(1996). RN [3] RP X-RAY CRYSTALLOGRAPHY (2.40 ANGSTROMS) IN COMPLEXES WITH NADP; METHOTREXATE RP AND TRIMETREXATE, AND CATALYTIC ACTIVITY. RX PubMed=18536013; DOI=10.1002/prot.22115; RA Schormann N., Senkovich O., Walker K., Wright D.L., Anderson A.C., RA Rosowsky A., Ananthan S., Shinkre B., Velu S., Chattopadhyay D.; RT "Structure-based approach to pharmacophore identification, in silico RT screening, and three-dimensional quantitative structure-activity RT relationship studies for inhibitors of Trypanosoma cruzi dihydrofolate RT reductase function."; RL Proteins 73:889-901(2008). RN [4] RP X-RAY CRYSTALLOGRAPHY (3.00 ANGSTROMS) IN COMPLEXES WITH NADP; UMP; RP METHOTREXATE AND TRIMETREXATE, AND SUBUNIT. RX PubMed=19564691; DOI=10.1107/s090744490901230x; RA Senkovich O., Schormann N., Chattopadhyay D.; RT "Structures of dihydrofolate reductase-thymidylate synthase of Trypanosoma RT cruzi in the folate-free state and in complex with two antifolate drugs, RT trimetrexate and methotrexate."; RL Acta Crystallogr. D 65:704-716(2009). RN [5] RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) IN COMPLEXES WITH NADP; UMP AND THE RP SYNTHETIC INHIBITORS C-448; CYC AND Q-8, AND CATALYTIC ACTIVITY. RX PubMed=19923744; DOI=10.1107/s1744309109041979; RA Chitnumsub P., Yuvaniyama J., Chahomchuen T., Vilaivan T., Yuthavong Y.; RT "Crystallization and preliminary crystallographic studies of dihydrofolate RT reductase-thymidylate synthase from Trypanosoma cruzi, the Chagas disease RT pathogen."; RL Acta Crystallogr. F 65:1175-1178(2009). CC -!- FUNCTION: Bifunctional enzyme. Involved in de novo dTMP biosynthesis. CC Key enzyme in folate metabolism. Catalyzes an essential reaction for de CC novo glycine and purine synthesis, DNA precursor synthesis, and for the CC conversion of dUMP to dTMP. CC -!- CATALYTIC ACTIVITY: CC Reaction=(6S)-5,6,7,8-tetrahydrofolate + NADP(+) = 7,8-dihydrofolate + CC H(+) + NADPH; Xref=Rhea:RHEA:15009, ChEBI:CHEBI:15378, CC ChEBI:CHEBI:57451, ChEBI:CHEBI:57453, ChEBI:CHEBI:57783, CC ChEBI:CHEBI:58349; EC=1.5.1.3; CC -!- CATALYTIC ACTIVITY: CC Reaction=(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP = 7,8- CC dihydrofolate + dTMP; Xref=Rhea:RHEA:12104, ChEBI:CHEBI:15636, CC ChEBI:CHEBI:57451, ChEBI:CHEBI:63528, ChEBI:CHEBI:246422; CC EC=2.1.1.45; CC -!- PATHWAY: Cofactor biosynthesis; tetrahydrofolate biosynthesis; 5,6,7,8- CC tetrahydrofolate from 7,8-dihydrofolate: step 1/1. CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:19564691}. CC -!- SIMILARITY: In the N-terminal section; belongs to the dihydrofolate CC reductase family. {ECO:0000305}. CC -!- SIMILARITY: In the C-terminal section; belongs to the thymidylate CC synthase family. {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; L22484; AAB49898.1; -; Genomic_DNA. DR RefSeq; XP_815327.1; XM_810234.1. DR PDB; 2H2Q; X-ray; 2.40 A; A/B=1-521. DR PDB; 3CL9; X-ray; 3.30 A; A=1-521. DR PDB; 3CLB; X-ray; 3.00 A; A/B/C/D=1-521. DR PDB; 3HBB; X-ray; 3.00 A; A/B/C/D=1-521. DR PDB; 3INV; X-ray; 2.37 A; A/B=1-521. DR PDB; 3IRM; X-ray; 2.10 A; A/B/C/D=1-521. DR PDB; 3IRN; X-ray; 2.60 A; A/B/C/D=1-521. DR PDB; 3IRO; X-ray; 2.80 A; A/B/C/D=1-521. DR PDBsum; 2H2Q; -. DR PDBsum; 3CL9; -. DR PDBsum; 3CLB; -. DR PDBsum; 3HBB; -. DR PDBsum; 3INV; -. DR PDBsum; 3IRM; -. DR PDBsum; 3IRN; -. DR PDBsum; 3IRO; -. DR AlphaFoldDB; Q27793; -. DR SMR; Q27793; -. DR BindingDB; Q27793; -. DR ChEMBL; CHEMBL1163130; -. DR DrugCentral; Q27793; -. DR GeneID; 3547029; -. DR KEGG; tcr:509153.90; -. DR VEuPathDB; TriTrypDB:BCY84_16439; -. DR VEuPathDB; TriTrypDB:C3747_84g65; -. DR VEuPathDB; TriTrypDB:C4B63_30g231; -. DR VEuPathDB; TriTrypDB:ECC02_006008; -. DR VEuPathDB; TriTrypDB:Tc_MARK_668; -. DR VEuPathDB; TriTrypDB:TcBrA4_0087940; -. DR VEuPathDB; TriTrypDB:TcCL_ESM02245; -. DR VEuPathDB; TriTrypDB:TcCLB.420533.9; -. DR VEuPathDB; TriTrypDB:TcCLB.509153.90; -. DR VEuPathDB; TriTrypDB:TcCLB.510303.320; -. DR VEuPathDB; TriTrypDB:TCDM_01885; -. DR VEuPathDB; TriTrypDB:TcG_01595; -. DR VEuPathDB; TriTrypDB:TCSYLVIO_001858; -. DR VEuPathDB; TriTrypDB:TcYC6_0045790; -. DR OMA; SIPWRVP; -. DR BRENDA; 1.5.1.3; 6524. DR BRENDA; 2.1.1.45; 6524. DR SABIO-RK; Q27793; -. DR UniPathway; UPA00077; UER00158. DR EvolutionaryTrace; Q27793; -. DR GO; GO:0004146; F:dihydrofolate reductase activity; IEA:UniProtKB-EC. DR GO; GO:0004799; F:thymidylate synthase activity; IEA:UniProtKB-EC. DR GO; GO:0006231; P:dTMP biosynthetic process; IEA:InterPro. DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW. DR GO; GO:0006730; P:one-carbon metabolic process; IEA:UniProtKB-KW. DR GO; GO:0046654; P:tetrahydrofolate biosynthetic process; IEA:UniProtKB-UniPathway. DR CDD; cd00209; DHFR; 1. DR CDD; cd00351; TS_Pyrimidine_HMase; 1. DR Gene3D; 3.40.430.10; Dihydrofolate Reductase, subunit A; 1. DR Gene3D; 3.30.572.10; Thymidylate synthase/dCMP hydroxymethylase domain; 1. DR HAMAP; MF_00008; Thymidy_synth_bact; 1. DR InterPro; IPR024072; DHFR-like_dom_sf. DR InterPro; IPR012262; DHFR-TS. DR InterPro; IPR017925; DHFR_CS. DR InterPro; IPR001796; DHFR_dom. DR InterPro; IPR045097; Thymidate_synth/dCMP_Mease. DR InterPro; IPR023451; Thymidate_synth/dCMP_Mease_dom. DR InterPro; IPR036926; Thymidate_synth/dCMP_Mease_sf. DR InterPro; IPR000398; Thymidylate_synthase. DR InterPro; IPR020940; Thymidylate_synthase_AS. DR NCBIfam; TIGR03284; thym_sym; 1. DR PANTHER; PTHR11548:SF2; THYMIDYLATE SYNTHASE; 1. DR PANTHER; PTHR11548; THYMIDYLATE SYNTHASE 1; 1. DR Pfam; PF00186; DHFR_1; 1. DR Pfam; PF00303; Thymidylat_synt; 1. DR PIRSF; PIRSF000389; DHFR-TS; 1. DR PRINTS; PR00108; THYMDSNTHASE. DR SUPFAM; SSF53597; Dihydrofolate reductase-like; 1. DR SUPFAM; SSF55831; Thymidylate synthase/dCMP hydroxymethylase; 1. DR PROSITE; PS00075; DHFR_1; 1. DR PROSITE; PS51330; DHFR_2; 1. DR PROSITE; PS00091; THYMIDYLATE_SYNTHASE; 1. PE 1: Evidence at protein level; KW 3D-structure; Methyltransferase; Multifunctional enzyme; NADP; KW Nucleotide biosynthesis; One-carbon metabolism; Oxidoreductase; KW Transferase. FT CHAIN 1..521 FT /note="Bifunctional dihydrofolate reductase-thymidylate FT synthase" FT /id="PRO_0000186354" FT DOMAIN 22..232 FT /note="DHFR" FT REGION 237..521 FT /note="Thymidylate synthase" FT ACT_SITE 403 FT /evidence="ECO:0000250" FT BINDING 26 FT /ligand="substrate" FT BINDING 28 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT BINDING 34..40 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT BINDING 48 FT /ligand="substrate" FT BINDING 78..80 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT BINDING 99..102 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT BINDING 154 FT /ligand="substrate" FT BINDING 155..162 FT /ligand="NADP(+)" FT /ligand_id="ChEBI:CHEBI:58349" FT BINDING 160 FT /ligand="substrate" FT BINDING 178 FT /ligand="substrate" FT BINDING 257 FT /ligand="dUMP" FT /ligand_id="ChEBI:CHEBI:246422" FT BINDING 404 FT /ligand="dUMP" FT /ligand_id="ChEBI:CHEBI:246422" FT BINDING 422..426 FT /ligand="dUMP" FT /ligand_id="ChEBI:CHEBI:246422" FT BINDING 434 FT /ligand="dUMP" FT /ligand_id="ChEBI:CHEBI:246422" FT BINDING 464..466 FT /ligand="dUMP" FT /ligand_id="ChEBI:CHEBI:246422" FT HELIX 2..4 FT /evidence="ECO:0007829|PDB:3IRM" FT TURN 10..13 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 14..16 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 23..30 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 33..39 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 46..57 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 60..63 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 70..77 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 78..82 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 86..88 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 94..99 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 105..110 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 112..114 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 115..121 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 122..124 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 125..130 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 132..139 FT /evidence="ECO:0007829|PDB:3IRM" FT TURN 142..147 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 148..153 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 157..163 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 168..171 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 172..180 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 188..190 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 199..201 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 206..211 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 218..221 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 224..232 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 235..250 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 252..255 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 261..273 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 275..277 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 283..285 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 289..300 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 307..310 FT /evidence="ECO:0007829|PDB:3IRM" FT TURN 311..313 FT /evidence="ECO:0007829|PDB:3IRM" FT TURN 319..321 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 323..328 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 343..349 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 357..359 FT /evidence="ECO:0007829|PDB:3CL9" FT HELIX 368..378 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 386..388 FT /evidence="ECO:0007829|PDB:3IRM" FT TURN 392..394 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 395..397 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 398..400 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 403..412 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 415..426 FT /evidence="ECO:0007829|PDB:3IRM" FT TURN 427..429 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 430..448 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 452..466 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 467..469 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 470..476 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 486..490 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 496..498 FT /evidence="ECO:0007829|PDB:3IRM" FT HELIX 501..503 FT /evidence="ECO:0007829|PDB:3IRM" FT STRAND 504..508 FT /evidence="ECO:0007829|PDB:3IRM" SQ SEQUENCE 521 AA; 58853 MW; DCB9F8782855810C CRC64; MSLFKIRMPE TVAEGTRLAL RAFSLVVAVD EHGGIGDGRS IPWNVPEDMK FFRDLTTKLR GKNVKPSPAK RNAVVMGRKT WDSIPPKFRP LPGRLNVVLS STLTTQHLLD GLPDEEKRNL HADSIVAVNG GLEQALRLLA SPNYTPSIET VYCIGGGSVY AEALRPPCVH LLQAIYRTTI RASESSCSVF FRVPESGTEA AAGIEWQRET ISEELTSANG NETKYYFEKL IPRNREEEQY LSLVDRIIRE GNVKHDRTGV GTLSIFGAQM RFSLRNNRLP LLTTKRVFWR GVCEELLWFL RGETYAKKLS DKGVHIWDDN GSRAFLDSRG LTEYEEMDLG PVYGFQWRHF GAAYTHHDAN YDGQGVDQIK AIVETLKTNP DDRRMLFTAW NPSALPRMAL PPCHLLAQFY VSNGELSCML YQRSCDMGLG VPFNIASYAL LTILIAKATG LRPGELVHTL GDAHVYSNHV EPCNEQLKRV PRAFPYLVFR REREFLEDYE EGDMEVIDYA PYPPISMKMA V //