THYX_THEMA
ID THYX_THEMA Reviewed; 220 AA.
AC Q9WYT0;
DT 11-JUL-2001, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1999, sequence version 1.
DT 03-AUG-2022, entry version 144.
DE RecName: Full=Flavin-dependent thymidylate synthase {ECO:0000303|PubMed:19370033, ECO:0000303|PubMed:23019356, ECO:0000303|PubMed:24563811, ECO:0000303|PubMed:27214228};
DE Short=FDTS {ECO:0000303|PubMed:19370033, ECO:0000303|PubMed:23019356, ECO:0000303|PubMed:24563811, ECO:0000303|PubMed:27214228};
DE EC=2.1.1.148 {ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:19370033};
DE AltName: Full=FAD-dependent thymidylate synthase {ECO:0000255|HAMAP-Rule:MF_01408};
DE AltName: Full=Thymidylate synthase ThyX {ECO:0000255|HAMAP-Rule:MF_01408};
DE Short=TS {ECO:0000255|HAMAP-Rule:MF_01408};
DE Short=TSase {ECO:0000255|HAMAP-Rule:MF_01408};
GN Name=thyX {ECO:0000303|PubMed:19370033};
GN Synonyms=thy1 {ECO:0000303|PubMed:12211025}; OrderedLocusNames=TM_0449;
OS Thermotoga maritima (strain ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826
OS / MSB8).
OC Bacteria; Thermotogae; Thermotogales; Thermotogaceae; Thermotoga.
OX NCBI_TaxID=243274;
RN [1]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8;
RX PubMed=10360571; DOI=10.1038/20601;
RA Nelson K.E., Clayton R.A., Gill S.R., Gwinn M.L., Dodson R.J., Haft D.H.,
RA Hickey E.K., Peterson J.D., Nelson W.C., Ketchum K.A., McDonald L.A.,
RA Utterback T.R., Malek J.A., Linher K.D., Garrett M.M., Stewart A.M.,
RA Cotton M.D., Pratt M.S., Phillips C.A., Richardson D.L., Heidelberg J.F.,
RA Sutton G.G., Fleischmann R.D., Eisen J.A., White O., Salzberg S.L.,
RA Smith H.O., Venter J.C., Fraser C.M.;
RT "Evidence for lateral gene transfer between Archaea and Bacteria from
RT genome sequence of Thermotoga maritima.";
RL Nature 399:323-329(1999).
RN [2]
RP X-RAY CRYSTALLOGRAPHY (2.25 ANGSTROMS) IN COMPLEX WITH FAD, COFACTOR, AND
RP SUBUNIT.
RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8;
RX PubMed=12211025; DOI=10.1002/prot.10202;
RA Kuhn P., Lesley S.A., Mathews I.I., Canaves J.M., Brinen L.S., Dai X.,
RA Deacon A.M., Elsliger M.-A., Eshaghi S., Floyd R., Godzik A., Grittini C.,
RA Grzechnik S.K., Guda C., Hodgson K.O., Jaroszewski L., Karlak C.,
RA Klock H.E., Koesema E., Kovarik J.M., Kreusch A.T., McMullan D.,
RA McPhillips T.M., Miller M.A., Miller M., Morse A., Moy K., Ouyang J.,
RA Robb A., Rodrigues K., Selby T.L., Spraggon G., Stevens R.C., Taylor S.S.,
RA van den Bedem H., Velasquez J., Vincent J., Wang X., West B., Wolf G.,
RA Wooley J., Wilson I.A.;
RT "Crystal structure of thy1, a thymidylate synthase complementing protein
RT from Thermotoga maritima at 2.25 A resolution.";
RL Proteins 49:142-145(2002).
RN [3] {ECO:0007744|PDB:1O24, ECO:0007744|PDB:1O25, ECO:0007744|PDB:1O26, ECO:0007744|PDB:1O27, ECO:0007744|PDB:1O28, ECO:0007744|PDB:1O29, ECO:0007744|PDB:1O2A, ECO:0007744|PDB:1O2B}
RP X-RAY CRYSTALLOGRAPHY (1.60 ANGSTROMS) OF APOENZYME AND COMPLEXES WITH FAD;
RP DUMP AND SUBSTRATE ANALOGS, FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND
RP SUBUNIT.
RC STRAIN=ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8;
RX PubMed=12791256; DOI=10.1016/s0969-2126(03)00097-2;
RA Mathews I.I., Deacon A.M., Canaves J.M., McMullan D., Lesley S.A.,
RA Agarwalla S., Kuhn P.;
RT "Functional analysis of substrate and cofactor complex structures of a
RT thymidylate synthase-complementing protein.";
RL Structure 11:677-690(2003).
RN [4] {ECO:0007744|PDB:3G4A, ECO:0007744|PDB:3G4C}
RP X-RAY CRYSTALLOGRAPHY (1.95 ANGSTROMS) OF MUTANTS ALA-88 AND CYS-88 IN
RP COMPLEX WITH FAD AND DUMP, FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, REACTION MECHANISM, AND MUTAGENESIS OF SER-88.
RX PubMed=19370033; DOI=10.1038/nature07973;
RA Koehn E.M., Fleischmann T., Conrad J.A., Palfey B.A., Lesley S.A.,
RA Mathews I.I., Kohen A.;
RT "An unusual mechanism of thymidylate biosynthesis in organisms containing
RT the thyX gene.";
RL Nature 458:919-923(2009).
RN [5] {ECO:0007744|PDB:3N0B, ECO:0007744|PDB:3N0C}
RP X-RAY CRYSTALLOGRAPHY (2.30 ANGSTROMS) IN COMPLEX WITH FAD AND DUMP.
RX PubMed=22477781; DOI=10.1107/s0021889810040409;
RA Berger M.A., Decker J.H., Mathews I.I.;
RT "Diffraction study of protein crystals grown in cryoloops and
RT micromounts.";
RL J. Appl. Crystallogr. 43:1513-1518(2010).
RN [6] {ECO:0007744|PDB:4GT9, ECO:0007744|PDB:4GTA, ECO:0007744|PDB:4GTB, ECO:0007744|PDB:4GTC, ECO:0007744|PDB:4GTD, ECO:0007744|PDB:4GTE, ECO:0007744|PDB:4GTF, ECO:0007744|PDB:4GTL}
RP X-RAY CRYSTALLOGRAPHY (1.39 ANGSTROMS) OF WILD-TYPE AND MUTANTS ALA-53;
RP ARG-144 AND LYS-174 IN COMPLEXES WITH FAD; DUMP AND SEVERAL FOLATE
RP DERIVATIVES, REACTION MECHANISM, AND MUTAGENESIS OF HIS-53; GLU-144 AND
RP ARG-174.
RX PubMed=23019356; DOI=10.1073/pnas.1206077109;
RA Koehn E.M., Perissinotti L.L., Moghram S., Prabhakar A., Lesley S.A.,
RA Mathews I.I., Kohen A.;
RT "Folate binding site of flavin-dependent thymidylate synthase.";
RL Proc. Natl. Acad. Sci. U.S.A. 109:15722-15727(2012).
RN [7] {ECO:0007744|PDB:4KAR, ECO:0007744|PDB:4KAS, ECO:0007744|PDB:4KAT}
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) OF MUTANT ASP-53 IN COMPLEXES WITH
RP FAD; FADH2 AND DUMP, AND SUBUNIT.
RX PubMed=24563811; DOI=10.4172/2157-2526.s12-004;
RA Mathews I.I.;
RT "Flavin-dependent thymidylate synthase as a drug target for deadly
RT microbes: mutational study and a strategy for inhibitor design.";
RL J. Bioterror. Biodef. 12:004-004(2013).
RN [8] {ECO:0007744|PDB:5IOQ, ECO:0007744|PDB:5IOR, ECO:0007744|PDB:5IOS, ECO:0007744|PDB:5IOT}
RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF WILD-TYPE AND MUTANTS ALA-90 AND
RP ALA-174 IN COMPLEXES WITH FAD; DUMP AND DEOXYURIDINE, REACTION MECHANISM,
RP ACTIVE SITE, AND MUTAGENESIS OF ARG-90 AND ARG-174.
RX PubMed=27214228; DOI=10.1021/acs.biochem.6b00510;
RA Stull F.W., Bernard S.M., Sapra A., Smith J.L., Zuiderweg E.R.,
RA Palfey B.A.;
RT "Deprotonations in the reaction of flavin-dependent thymidylate synthase.";
RL Biochemistry 55:3261-3269(2016).
RN [9] {ECO:0007744|PDB:7NDW, ECO:0007744|PDB:7NDZ}
RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) IN COMPLEXES WITH A FLAVIN
RP CARBINOLAMINE AND FAD, FUNCTION, CATALYTIC ACTIVITY, AND REACTION
RP MECHANISM.
RX PubMed=34315871; DOI=10.1038/s41467-021-24756-8;
RA Bou-Nader C., Stull F.W., Pecqueur L., Simon P., Guerineau V., Royant A.,
RA Fontecave M., Lombard M., Palfey B.A., Hamdane D.;
RT "An enzymatic activation of formaldehyde for nucleotide methylation.";
RL Nat. Commun. 12:4542-4542(2021).
CC -!- FUNCTION: Catalyzes the reductive methylation of 2'-deoxyuridine-5'-
CC monophosphate (dUMP or deoxyuridylate) to 2'-deoxythymidine-5'-
CC monophosphate (dTMP or deoxythymidylate) while utilizing 5,10-
CC methylenetetrahydrofolate (mTHF) as the methylene donor, and NAD(P)H
CC and FADH(2) as the reductant. This reaction is a critical step in DNA
CC biosynthesis (PubMed:12791256, PubMed:19370033). Can also use
CC formaldehyde instead of mTHF as a direct methylene donor for dTMP
CC synthesis. However, the tighter binding of ThyX to mTHF (KD of 4 uM)
CC compared to formaldehyde (KD of 20 mM) confirms that methylene
CC tetrahydrofolate acts as the biological carbon donor for ThyX, serving
CC as a formaldehyde carrier/transporter and thus avoiding genotoxic
CC effects (PubMed:34315871). {ECO:0000269|PubMed:12791256,
CC ECO:0000269|PubMed:19370033, ECO:0000269|PubMed:34315871}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate + dUMP + H(+) +
CC NADPH = (6S)-5,6,7,8-tetrahydrofolate + dTMP + NADP(+);
CC Xref=Rhea:RHEA:29043, ChEBI:CHEBI:15378, ChEBI:CHEBI:15636,
CC ChEBI:CHEBI:57453, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:63528, ChEBI:CHEBI:246422; EC=2.1.1.148;
CC Evidence={ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:19370033};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=dUMP + formaldehyde + H(+) + NADPH = dTMP + H2O + NADP(+);
CC Xref=Rhea:RHEA:68268, ChEBI:CHEBI:15377, ChEBI:CHEBI:15378,
CC ChEBI:CHEBI:16842, ChEBI:CHEBI:57783, ChEBI:CHEBI:58349,
CC ChEBI:CHEBI:63528, ChEBI:CHEBI:246422;
CC Evidence={ECO:0000269|PubMed:34315871};
CC -!- COFACTOR:
CC Name=FAD; Xref=ChEBI:CHEBI:57692;
CC Evidence={ECO:0000269|PubMed:12791256, ECO:0000305|PubMed:12211025};
CC Note=Binds 4 FAD per tetramer. Each FAD binding site is formed by three
CC monomers. {ECO:0000269|PubMed:12211025, ECO:0000269|PubMed:12791256};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=30 uM for dUMP (at 65 degrees Celsius)
CC {ECO:0000269|PubMed:19370033};
CC Note=kcat is 1.2 sec(-1) (at 65 degrees Celsius).
CC {ECO:0000269|PubMed:19370033};
CC -!- PATHWAY: Pyrimidine metabolism; dTTP biosynthesis. {ECO:0000255|HAMAP-
CC Rule:MF_01408}.
CC -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:12211025,
CC ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:24563811}.
CC -!- MISCELLANEOUS: Reaction mechanism involved a direct methylene transfer
CC from mTHF to dUMP (PubMed:23019356). FDTS ionizes N3 of dUMP using the
CC active-site Arg-174, providing a new mechanism for dUMP activation. The
CC phosphate of dUMP is crucial for flavin oxidation, suggesting that it
CC acts as the base that deprotonates C5 of the dUMP-methylene adduct
CC (PubMed:27214228). A later study showed that FAD is first reduced by
CC NADPH. Then, the reduced flavin, FADH(-), reacts with mTHF to form a
CC carbinolamine flavin, which acts as the genuine methylene donor. The
CC flavin carbinolamine can be obtained directly via a CH2O-shunt reaction
CC consisting of a reaction of FADH(-) with free CH2O. Methylene transfer
CC from FAD to dUMP is initiated by an SN2 reaction of activated dUMP and
CC the flavin carbinolamine, leading to water elimination and formation of
CC a transient FAD-CH2-dUMP adduct (PubMed:34315871).
CC {ECO:0000269|PubMed:23019356, ECO:0000269|PubMed:27214228,
CC ECO:0000269|PubMed:34315871}.
CC -!- SIMILARITY: Belongs to the thymidylate synthase ThyX family.
CC {ECO:0000255|HAMAP-Rule:MF_01408}.
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DR EMBL; AE000512; AAD35532.1; -; Genomic_DNA.
DR PIR; B72375; B72375.
DR RefSeq; NP_228259.1; NC_000853.1.
DR RefSeq; WP_004081517.1; NZ_CP011107.1.
DR PDB; 1KQ4; X-ray; 2.25 A; A/B/C/D=1-220.
DR PDB; 1O24; X-ray; 2.00 A; A/B/C/D=1-220.
DR PDB; 1O25; X-ray; 2.40 A; A/B/C/D=1-220.
DR PDB; 1O26; X-ray; 1.60 A; A/B/C/D=1-220.
DR PDB; 1O27; X-ray; 2.30 A; A/B/C/D=1-220.
DR PDB; 1O28; X-ray; 2.10 A; A/B/C/D=1-220.
DR PDB; 1O29; X-ray; 2.00 A; A/B/C/D=1-220.
DR PDB; 1O2A; X-ray; 1.80 A; A/B/C/D=1-220.
DR PDB; 1O2B; X-ray; 2.45 A; A/B/C/D=1-220.
DR PDB; 3G4A; X-ray; 1.95 A; A/B/C/D=1-220.
DR PDB; 3G4C; X-ray; 2.05 A; A/B/C/D=1-220.
DR PDB; 3N0B; X-ray; 2.30 A; A/B/C/D=1-220.
DR PDB; 3N0C; X-ray; 2.30 A; A/B/C/D=1-220.
DR PDB; 4GT9; X-ray; 1.39 A; A=1-220.
DR PDB; 4GTA; X-ray; 1.50 A; A=1-220.
DR PDB; 4GTB; X-ray; 1.70 A; A=1-220.
DR PDB; 4GTC; X-ray; 1.97 A; A/B/C/D=1-220.
DR PDB; 4GTD; X-ray; 1.76 A; A/B/C/D=1-220.
DR PDB; 4GTE; X-ray; 1.89 A; A/B/C/D=1-220.
DR PDB; 4GTF; X-ray; 1.77 A; A=1-220.
DR PDB; 4GTL; X-ray; 2.17 A; A/B/C/D=1-220.
DR PDB; 4KAR; X-ray; 2.03 A; A/B/C/D=1-220.
DR PDB; 4KAS; X-ray; 1.85 A; A/B/C/D=1-220.
DR PDB; 4KAT; X-ray; 2.14 A; A/B/C/D=1-220.
DR PDB; 5CHP; X-ray; 1.70 A; A=1-220.
DR PDB; 5IOQ; X-ray; 1.93 A; A/B/C/D=1-220.
DR PDB; 5IOR; X-ray; 1.95 A; A=1-220.
DR PDB; 5IOS; X-ray; 1.90 A; A/B/C/D=1-220.
DR PDB; 5IOT; X-ray; 2.00 A; A/B/C/D=1-220.
DR PDB; 5JFE; X-ray; 2.03 A; A=1-220.
DR PDB; 7NDW; X-ray; 2.00 A; A/B/C/D=1-220.
DR PDB; 7NDZ; X-ray; 2.70 A; A/B/C/D=1-220.
DR PDBsum; 1KQ4; -.
DR PDBsum; 1O24; -.
DR PDBsum; 1O25; -.
DR PDBsum; 1O26; -.
DR PDBsum; 1O27; -.
DR PDBsum; 1O28; -.
DR PDBsum; 1O29; -.
DR PDBsum; 1O2A; -.
DR PDBsum; 1O2B; -.
DR PDBsum; 3G4A; -.
DR PDBsum; 3G4C; -.
DR PDBsum; 3N0B; -.
DR PDBsum; 3N0C; -.
DR PDBsum; 4GT9; -.
DR PDBsum; 4GTA; -.
DR PDBsum; 4GTB; -.
DR PDBsum; 4GTC; -.
DR PDBsum; 4GTD; -.
DR PDBsum; 4GTE; -.
DR PDBsum; 4GTF; -.
DR PDBsum; 4GTL; -.
DR PDBsum; 4KAR; -.
DR PDBsum; 4KAS; -.
DR PDBsum; 4KAT; -.
DR PDBsum; 5CHP; -.
DR PDBsum; 5IOQ; -.
DR PDBsum; 5IOR; -.
DR PDBsum; 5IOS; -.
DR PDBsum; 5IOT; -.
DR PDBsum; 5JFE; -.
DR PDBsum; 7NDW; -.
DR PDBsum; 7NDZ; -.
DR AlphaFoldDB; Q9WYT0; -.
DR SMR; Q9WYT0; -.
DR DIP; DIP-60076N; -.
DR STRING; 243274.THEMA_02480; -.
DR DrugBank; DB01903; 5-Bromo-2'-deoxyuridine 5'-(dihydrogen phosphate).
DR DrugBank; DB03761; 5-fluoro-2'-deoxyuridine-5'-monophosphate.
DR DrugBank; DB03800; Deoxyuridine monophosphate.
DR DrugBank; DB03147; Flavin adenine dinucleotide.
DR EnsemblBacteria; AAD35532; AAD35532; TM_0449.
DR KEGG; tma:TM0449; -.
DR eggNOG; COG1351; Bacteria.
DR InParanoid; Q9WYT0; -.
DR OMA; EWARLMP; -.
DR OrthoDB; 896350at2; -.
DR BioCyc; MetaCyc:MON-15758; -.
DR BRENDA; 2.1.1.148; 6331.
DR SABIO-RK; Q9WYT0; -.
DR UniPathway; UPA00575; -.
DR EvolutionaryTrace; Q9WYT0; -.
DR Proteomes; UP000008183; Chromosome.
DR GO; GO:0050660; F:flavin adenine dinucleotide binding; IBA:GO_Central.
DR GO; GO:0070402; F:NADPH binding; IBA:GO_Central.
DR GO; GO:0050797; F:thymidylate synthase (FAD) activity; IBA:GO_Central.
DR GO; GO:0004799; F:thymidylate synthase activity; IBA:GO_Central.
DR GO; GO:0006231; P:dTMP biosynthetic process; IBA:GO_Central.
DR GO; GO:0006235; P:dTTP biosynthetic process; IEA:UniProtKB-UniPathway.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR Gene3D; 3.30.1360.170; -; 1.
DR HAMAP; MF_01408; ThyX; 1.
DR InterPro; IPR003669; Thymidylate_synthase_ThyX.
DR InterPro; IPR036098; Thymidylate_synthase_ThyX_sf.
DR PANTHER; PTHR34934; PTHR34934; 1.
DR Pfam; PF02511; Thy1; 1.
DR SUPFAM; SSF69796; SSF69796; 1.
DR TIGRFAMs; TIGR02170; thyX; 1.
DR PROSITE; PS51331; THYX; 1.
PE 1: Evidence at protein level;
KW 3D-structure; FAD; Flavoprotein; Methyltransferase; NAD; NADP;
KW Nucleotide biosynthesis; Reference proteome; Transferase.
FT CHAIN 1..220
FT /note="Flavin-dependent thymidylate synthase"
FT /id="PRO_0000175579"
FT DOMAIN 1..208
FT /note="ThyX"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00661"
FT MOTIF 78..88
FT /note="ThyX motif"
FT /evidence="ECO:0000255|HAMAP-Rule:MF_01408"
FT ACT_SITE 174
FT /note="Involved in ionization of N3 of dUMP, leading to its
FT activation"
FT /evidence="ECO:0000269|PubMed:27214228"
FT BINDING 55
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /ligand_note="ligand shared between neighboring subunits"
FT /evidence="ECO:0000269|PubMed:12791256,
FT ECO:0000269|PubMed:19370033, ECO:0000269|PubMed:22477781,
FT ECO:0000269|PubMed:23019356, ECO:0000269|PubMed:24563811,
FT ECO:0000269|PubMed:27214228, ECO:0000269|PubMed:34315871,
FT ECO:0007744|PDB:1O26, ECO:0007744|PDB:3G4A,
FT ECO:0007744|PDB:3N0B, ECO:0007744|PDB:4GTD,
FT ECO:0007744|PDB:4KAS, ECO:0007744|PDB:7NDW"
FT BINDING 75..78
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /evidence="ECO:0000269|PubMed:12791256,
FT ECO:0000269|PubMed:19370033, ECO:0000269|PubMed:22477781,
FT ECO:0000269|PubMed:23019356, ECO:0000269|PubMed:24563811,
FT ECO:0000269|PubMed:27214228, ECO:0007744|PDB:1O26,
FT ECO:0007744|PDB:3G4A, ECO:0007744|PDB:3N0B,
FT ECO:0007744|PDB:4GT9, ECO:0007744|PDB:4KAS"
FT BINDING 78..81
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /ligand_note="ligand shared between neighboring subunits"
FT /evidence="ECO:0000269|PubMed:12211025,
FT ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:19370033,
FT ECO:0000269|PubMed:22477781, ECO:0000269|PubMed:23019356,
FT ECO:0000269|PubMed:24563811, ECO:0000269|PubMed:27214228,
FT ECO:0000269|PubMed:34315871, ECO:0007744|PDB:1KQ4,
FT ECO:0007744|PDB:1O26, ECO:0007744|PDB:3G4A,
FT ECO:0007744|PDB:3N0B, ECO:0007744|PDB:4GTD,
FT ECO:0007744|PDB:4KAS, ECO:0007744|PDB:7NDW"
FT BINDING 86..90
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000269|PubMed:12791256,
FT ECO:0000269|PubMed:19370033, ECO:0000269|PubMed:22477781,
FT ECO:0000269|PubMed:23019356, ECO:0000269|PubMed:24563811,
FT ECO:0000269|PubMed:27214228, ECO:0007744|PDB:1O26,
FT ECO:0007744|PDB:3G4A, ECO:0007744|PDB:3N0B,
FT ECO:0007744|PDB:4GT9, ECO:0007744|PDB:4KAS"
FT BINDING 86
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /ligand_note="ligand shared between neighboring subunits"
FT /evidence="ECO:0000269|PubMed:12211025,
FT ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:19370033,
FT ECO:0000269|PubMed:22477781, ECO:0000269|PubMed:23019356,
FT ECO:0000269|PubMed:24563811, ECO:0000269|PubMed:27214228,
FT ECO:0000269|PubMed:34315871, ECO:0007744|PDB:1KQ4,
FT ECO:0007744|PDB:1O26, ECO:0007744|PDB:3G4A,
FT ECO:0007744|PDB:3N0B, ECO:0007744|PDB:4GTD,
FT ECO:0007744|PDB:4KAS, ECO:0007744|PDB:7NDW"
FT BINDING 147
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /note="in other chain"
FT /evidence="ECO:0000269|PubMed:12791256,
FT ECO:0000269|PubMed:19370033, ECO:0000269|PubMed:22477781,
FT ECO:0000269|PubMed:23019356, ECO:0000269|PubMed:24563811,
FT ECO:0000269|PubMed:27214228, ECO:0007744|PDB:1O26,
FT ECO:0007744|PDB:3G4A, ECO:0007744|PDB:3N0B,
FT ECO:0007744|PDB:4GT9, ECO:0007744|PDB:4KAS"
FT BINDING 163..165
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /ligand_note="ligand shared between neighboring subunits"
FT /evidence="ECO:0000269|PubMed:12211025,
FT ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:19370033,
FT ECO:0000269|PubMed:22477781, ECO:0000269|PubMed:23019356,
FT ECO:0000269|PubMed:24563811, ECO:0000269|PubMed:27214228,
FT ECO:0000269|PubMed:34315871, ECO:0007744|PDB:1KQ4,
FT ECO:0007744|PDB:1O26, ECO:0007744|PDB:3G4A,
FT ECO:0007744|PDB:3N0B, ECO:0007744|PDB:4GTD,
FT ECO:0007744|PDB:4KAS, ECO:0007744|PDB:7NDW"
FT BINDING 169
FT /ligand="FAD"
FT /ligand_id="ChEBI:CHEBI:57692"
FT /ligand_note="ligand shared between neighboring subunits"
FT /evidence="ECO:0000269|PubMed:12211025,
FT ECO:0000269|PubMed:12791256, ECO:0000269|PubMed:19370033,
FT ECO:0000269|PubMed:22477781, ECO:0000269|PubMed:23019356,
FT ECO:0000269|PubMed:24563811, ECO:0000269|PubMed:27214228,
FT ECO:0000269|PubMed:34315871, ECO:0007744|PDB:1KQ4,
FT ECO:0007744|PDB:1O26, ECO:0007744|PDB:3G4A,
FT ECO:0007744|PDB:3N0B, ECO:0007744|PDB:4GTD,
FT ECO:0007744|PDB:4KAS, ECO:0007744|PDB:7NDW"
FT BINDING 174
FT /ligand="dUMP"
FT /ligand_id="ChEBI:CHEBI:246422"
FT /ligand_note="ligand shared between dimeric partners"
FT /evidence="ECO:0000269|PubMed:12791256,
FT ECO:0000269|PubMed:19370033, ECO:0000269|PubMed:22477781,
FT ECO:0000269|PubMed:23019356, ECO:0000269|PubMed:24563811,
FT ECO:0000269|PubMed:27214228, ECO:0007744|PDB:1O26,
FT ECO:0007744|PDB:3G4A, ECO:0007744|PDB:3N0B,
FT ECO:0007744|PDB:4GT9, ECO:0007744|PDB:4KAS"
FT MUTAGEN 53
FT /note="H->A: Shows 1.39% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:23019356"
FT MUTAGEN 88
FT /note="S->A,C: Still catalytically active although shows a
FT large decrease in activity."
FT /evidence="ECO:0000269|PubMed:19370033"
FT MUTAGEN 90
FT /note="R->A: Binds dUMP 670-fold weaker than wild-type."
FT /evidence="ECO:0000269|PubMed:27214228"
FT MUTAGEN 144
FT /note="E->A: Shows 0.113% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:23019356"
FT MUTAGEN 144
FT /note="E->R: Shows 0.016% of wild-type activity."
FT /evidence="ECO:0000269|PubMed:23019356"
FT MUTAGEN 174
FT /note="R->A: Still catalytically active although only shows
FT 0.0008% of wild-type activity. Binds dUMP 7300-fold weaker
FT than wild-type."
FT /evidence="ECO:0000269|PubMed:23019356,
FT ECO:0000269|PubMed:27214228"
FT MUTAGEN 174
FT /note="R->K: Loss of catalytic activity."
FT /evidence="ECO:0000269|PubMed:23019356"
FT STRAND 2..5
FT /evidence="ECO:0007829|PDB:4GT9"
FT TURN 6..8
FT /evidence="ECO:0007829|PDB:4GT9"
FT STRAND 9..17
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 20..28
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 29..31
FT /evidence="ECO:0007829|PDB:1O26"
FT HELIX 39..51
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 55..59
FT /evidence="ECO:0007829|PDB:4GT9"
FT STRAND 61..69
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 70..76
FT /evidence="ECO:0007829|PDB:4GT9"
FT STRAND 81..86
FT /evidence="ECO:0007829|PDB:4GT9"
FT TURN 89..91
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 103..106
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 115..138
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 143..146
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 147..149
FT /evidence="ECO:0007829|PDB:4GT9"
FT STRAND 154..163
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 164..174
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 181..197
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 199..208
FT /evidence="ECO:0007829|PDB:4GT9"
FT HELIX 214..216
FT /evidence="ECO:0007829|PDB:1O26"
SQ SEQUENCE 220 AA; 26004 MW; E3B9712014185907 CRC64;
MKIDILDKGF VELVDVMGND LSAVRAARVS FDMGLKDEER DRHLIEYLMK HGHETPFEHI
VFTFHVKAPI FVARQWFRHR IASYNELSGR YSKLSYEFYI PSPERLEGYK TTIPPERVTE
KISEIVDKAY RTYLELIESG VPREVARIVL PLNLYTRFFW TVNARSLMNF LNLRADSHAQ
WEIQQYALAI ARIFKEKCPW TFEAFLKYAY KGDILKEVQV
